Note: Descriptions are shown in the official language in which they were submitted.
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COMPOUNDS WITH GROWTH HORMONE RELEASING PROPERTIES
FIELD OF INVENTION
... 5
The present invention relates to novel compounds, compositions containing
them, and their
use for treating medical disorders resulting from a deficiency in growth
hormone.
BACKGROUND OF THE INVENTION
Growth hormone is a hormone which stimulates growth of all tissues capable of
growing. In
addition, growth hormone is known to have a number of effects on metabolic
processes, e.g.,
stimulation of protein synthesis and free fatty acid mobilisation and to cause
a switch in energy
metabolism from carbohydrate to fatty acid metabolism. Deficiency in growth
hormone can
~5 result in a number of severe medical disorders, e.g., dwarfism.
Growth hormone is released from the pituitary. The release is under tight
control of a number
of hormones and neurotransmitters either directly or indirectly. Growth
hormone release can be
stimulated by growth hormone releasing hormone (GHRH) and inhibited by
somatostatin. In
both cases the hormones are released from the hypothalamus but their action is
mediated
primarily via specfic receptors located in the pituitary. Other compounds
which stimulate the
release of growth hormone from the pituitary have also been described. For
example arginine,
L-3,4-dihydroxyphenylalanine (L-Dopa), glucagon, vasopressin, PACAP (pituitary
adenylyl
cydase activating peptide), muscarinic receptor agonists and a synthetic
hexapeptide, GHRP
(growth hormone releasing peptide) release endogenous growth hormone either by
a direct
effect on the pituitary or by affecting the release of GHRH and/or
somatostatin from the
hypothalamus.
In disorders or conditions where increased levels of growth hormone is
desired, the protein
nature of growth hormone makes anything but parenteral administration non-
viable. Further-
more, other directly acting natural secretagogues, e.g., GHRH and PACAP, are
longer poly-
peptides for which reason parenteral administration is prefer-ed.
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The composition of growth hormone releasing compounds is important for their
growth hor-
mone releasing potency as well as their bioavailability. It is therefore an
object of the present
invention to provide novel compounds with growth hormone releasing properties.
Moreover, it
is an object to provide novel growth hormone releasing compounds (growth
hormone secre-
tagogues) which are specific andlor selective and have no or substantially no
side-effects, such
as e.g. release of t-H, FSH, TSH, ACTH, vasopressin, oxytocin, cortisol andlor
prolactin. It is
also an object to provide compounds which have good oral bioavailabifity.
2
The use of certain compounds for increasing the levels of growth hormone in
mammals has
previously been proposed, e.g. in EP 18 072, EP 83 864, WO 8302272, WO
8907110, WO
8901711, WO 8910933, WO 8809780, WO 9118016, WO 9201711, WO 9304081, WO
9413696, WO 9517423, WO 9514666, WO 9615148, WO 9622997, WO 9635713, WO
5 9700894, WO 9722620, WO 9723508, WO 9740023, and WO 9810653.
WO 00/01726 PCTIDK99/00368
SUMMARY OF THE INVENTION
In accordance with the present invention there is provided novel compounds
which act directly
on the pituitary cells under normal experimental conditions in vitro to
release growth hormone
therefrom.
These growth hormone releasing compounds can be utilized in vitro as unique
research tools
for understanding, inter alia, how growth hormone secretion is regulated at
the pituitary level.
Moreover, the growth hormone releasing compounds of the present invention can
also be
administered in vivo to increase endogenous growth hormone release.
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DESCRIPTION OF THE INVENTION
Accordingly, the present invention relates to a compound of the general
formula I
., 5
O aC' R2 O
Ra~N~ CR6R7~---~-~,M~(CHR5)d c N N
)f ~ ~ L
R' O
formula 1
wherein
R' is hydrogen or C,.~-alkyl;
R2 is hydrogen or C,.~-alkyl;
L is
R9 Rio
R Rio
r
4 s
~N ~q , ~)S -N
R4 ( Q or
( t
R'2 R~~ _ R~2 R~~
wherein R~ is hydrogen or C,.~ alkyl;
pis0orl;
q, s, t, a are independently from each other 0, 1, 2, 3 or 4;
ris0or1;
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the sum q + r + s + t + a is 0, 1, 2, 3, or 4;
R9, R'°, R", and R'Z are independently from each other hydrogen or C,.6
alkyl;
Q is >N-R'3 or
Rya
'~~ T .
0
wherein o is 0, 1 or 2;
T is -N(R'S)(R'6) or hydroxyl;
R", R'S, and R'6 are independently from each other hydrogen or C,_6 alkyl;
R" is hydrogen, aryl or hetaryl;
G is -O-(CHZ)k-R",
R,7 R,~ R,e
___~~ R, s ' ,
Rz, ~ R~9 , N
Ro R" R,~ R,e
R,a
R,s N \~~~.~ ~ /
g or g ,
N H
H
wherein R", R'°, R'9, R~° and RZ' independently from each other
are hydrogen, halogen, aryl,
hetaryl, C,~-alkyl or C,.~-alkoxy;
k is 0, 1 or 2;
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J is -O-(CH2),-Rte,
. Rzz Rzz R2z Rz3
/ Rz3
Rz3 ___~~.~~-- /
w w ,
Rzs w I R2a , N
R25 22 22 23
R2z R22 R R23
/ Rz3 N \1~'~ \ ~
~N ~ , S or S ,
H
H
PCT/DK99I00368
5 wherein R2z, Rz3, Rza, R2s and R2g independently from each other are
hydrogen, halogen, aryl,
hetaryl, C,.~-alkyl or C,~-alkoxy;
I is 0, 1 or 2;
a is 0, 1, or 2;
b is 0, 1, or 2;
c is 0, 1, or 2;
dis0or1;
a is 0, 1, 2, or 3;
fis0or1;
RS is hydrogen or C,.~-alkyl optionally substituted with one or more hydroxyl,
aryl or hetaryl;
R6 and R' are independently from each other hydrogen or C,.~-alkyl, optionally
substituted
- with one or more halogen, amino, hydroxyl, aryl, or hetaryl;
RB is hydrogen or C,~-alkyl, optionally substituted with one or more halogen,
amino, hy-
droxyl, aryl, or hetaryl;
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PCT/DK99/00368
R6 and R' or R6 and R° or R' and R8 may optionally form -(CHZ); U-
(CHZ)~ , wherein i and j
independently from each other are 1, 2 or 3 and U is -O-, -S-, or a valence
bond;
M is arylene, hetarylene, -O-, -S- or -CRZ'=CRIB-;
RZ' and RZB are independently from each other hydrogen or C,.e-alkyl,
optionally substituted
with one or more aryl or hetaryl;
or a pharmaceutically acceptable salt thereof.
Moreover, the compounds of formula i may comprise any optical isomers thereof,
in the form
of separated, pure or partially purified optical isomers or racemic mixtures
thereof. Whenever
one or more chiral carbon atoms are present such chiral center or centers may
be in the R-
andlor S-configuration, or a mixture of R and S.
Furthermore, the compounds of formula I may have one or more carbon-carbon
double
bonds with the possibility of geometric isomeri, and it is intended that
possible stereoisomers
(E or Z isomers) are included in the scope of the invention, unless a special
geometric
isomer is specified.
In one embodiment of the compound of formula I R' is C,~-alkyl, such as C,~-
alkyl, in
particular methyl. In a second embodiment R' is hydrogen.
In a further embodiment of the compound of formula l RZ is C,~-alkyl, such as
C,.~-alkyl, in
particular methyl.
in a still further embodiment of the compound of formula I L is
R9
I , R' o
w N a , ~)
a
R " .Q
R'Z'R~ ~
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wherein R° is hydrogen or C,.~ alkyl;
pis0or1;
q, s, t, a are independently from each other 0, 1, 2, 3 or 4;
PCTIDK99100368
ris0or1;
thesumq+r+s+t+uis0,1,2,3,or4;
R9, R'°, R", and R'2 are independently from each other hydrogen or C,.~
alkyl;
Q is >N-R'3 or
'/~T
wherein o is 0, 1 or 2;
T is -N(R'S)(R'6) or hydroxyl;
R", R'S, and R's are independently from each other hydrogen or C,.~ alkyl;
R" is hydrogen, aryl or hetaryl. In one embodiment R° is hydrogen. In a
second embodiment
R' is C,~-alkyl, such as C,.'-alkyl, in particular methyl. In a third
embodiment p is 0. In a fur
ther embodiment q is 0. fn a still further embodiment q is 1. In a further
embodiment s is 0. In
a still further embodiment s is 1. In a further embodiment t is 0. In a still
further embodiment t
is 1. In a further embodiment a is 0. In a still further embodiment a is 1. In
a further embodi-
ment r is 0. In a still further embodiment r is 1. In a further embodiment R9
is hydrogen. In a
still further embodiment Re is C,.~-alkyl, such as C,.; alkyl, in particular
methyl. In a further
embodiment R'° is hydrogen. In a still further embodiment R'° is
C,.~-alkyl, such as C,.°-alkyl,
in particular methyl. In a further embodiment R" is hydrogen. In a still
further embodiment
R" is C,~-alkyl, such as C,~-alkyl, in particular methyl. In a further
embodiment R'2 is hydro-
gen. In a still further embodiment R'2 is C,.~-alkyl, such as C,.,-alkyl, in
particular methyl. In a
further embodiment Q is >N-R". In a still further embodiment R" is hydrogen.
In a further
embodiment R'3 is C,~-alkyl, such as C,.'-alkyl, in particular methyl. In a
still further embodi-
ment Q is
Rya
. - ,~~T
In a still further embodiment R'° is hetaryl, in particular thiazolyl.
In a further embodiment R'°
is hydrogen. In a still further embodiment o is 0. In a further embodiment o
is 1. In a still fur-
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ther embodiment T is hydroxyl. In a further embodiment T is -N(R'S)(R'6). In a
still further
embodiment R'S is C,.~-alkyl, such as C,~ alkyl, in particular methyl. In a
further embodiment
R'6 is C,.~-alkyl, such as C,.,-alkyl, in particular methyl.
In a further embodiment of the compound of formula I L is
R~o
q s
-N Q
z
R»
wherein q, s, t, a independently from each other are 0, 1, 2, 3 or 4;
ris0or1;
the sum q + r + s + t + a is 0, 1, 2, 3, or 4;
Re, R'°, R", and R'2 are independently from each other hydrogen or C,.~
alkyl;
Q is >N-R'3 or
R, a
'~~ T
0
wherein o is 0, 1 or 2;
T is -N(R'S)(R'6) or hydroxyl;
R'3, R'S, and R'6 are independently from each other hydrogen or C,.~ alkyl;
R'° is hydrogen, aryl or hetaryl. In one embodiment q is 0. In a second
embodiment q is 1. 1n
a third embodiment s is 0. In a further embodiment s is 1. In a still further
embodiment t is 0.
In a further embodiment t is 1. In a still further embodiment a is 0. In a
further embodiment a
is 1. In a still further embodiment r is 0. In a further embodiment r is 1. In
a still further em-
bodiment R9 is hydrogen. In a still further embodiment R9 is C,~-alkyl, such
as C,.,-alkyl, in
particular methyl. In a further embodiment R'° is hydrogen. In a still
further embodiment R'°
is C,~-alkyl, such as C,.~-alkyl, in particular methyl. 1n a further
embodiment R" is hydrogen.
In a still further embodiment R" is C,~-alkyl, such as C,.,-alkyl, in
particular methyl. In a fur-
ther embodiment R'2 is hydrogen. In a still further embodiment R'2 is C,_6-
alkyl, such as C,.~
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alkyl, in particular methyl. In a further embodiment Q is >N-R" . In a still
further embodiment
R'3 is hydrogen. In a still further embodiment R'3 is C,.~-alkyl, such as C,.~-
alkyl, in particular
methyl. In a further embodiment Q is
R, a
' '/~ T
t o
In a still further embodiment R" is hetaryl, in particular thiazolyl. In a
further embodiment R'°
is hydrogen. In a still further embodiment o is 0. In a further embodiment o
is 1. In a still fur-
ther embodiment T is hydroxyl. In a further embodiment T is -N(R'S)(R'6). In a
still further
embodiment R'S is C,_6-alkyl, such as C,.e-alkyl, in particular methyl. In a
further embodiment
R'6 is C,.~-alkyl, such as C,~-alkyl, in particular methyl.
In the compound of the above formula I L is preferably 4-hydroxy-4-(2-
thienyl)piperidino, (3-
hydroxycyclohexyl)amino, 4-{N,N-dimethylamino)piperidino, N-methyl-N-(1-
methyipiperidin-
4-yl)amino), 4-((N,N-dirnethylamino)methyl)piperidino, 4-methyfpiperazino,
(2,2,6,6-tetra-
methylpiperidine-4-yl)amino, 4-hydroxypiperidino, (3S)-3-((N,N-
dimethylamino)methyl}-
piperidino, (2S)-2-((N,N-dimethylamino)methyl)pyrrolidino,
In a still further embodiment of the compound of formula I G is
R"
R'°
/ R,7 R,e
RZ, ~~. I R,a /
Rz° w w
Or
2o wherein R", R'e, R's, R~° and RZ' independently from each other are
hydrogen, halogen, aryl,
hetaryl, C,~-alkyl or C,.~-alkoxy. In one embodiment R" is hydrogen. In a
second embodiment
R'e is hydrogen. In a third embodiment R'9 is hydrogen. In a further
embodiment R'9 is aryl, in
particular phenyl. In a still further embodiment R~° is hydrogen. In a
further embodiment RZ' is
hydrogen. In the compound of the above formula I G is preferably 2-naphthyl or
biphenyl-4-yl.
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In a further embodiment of the compound of formula I J is
R22
R23
R
\\\~ R23
R26 w R2a
R25
5 wherein Rzz, R23, R2°, Rzs and Rzs independently from each other are
hydrogen, halogen, aryl,
hetaryl, C,~-alkyl or C,.~-alkoxy. in one embodiment Rzz is hydrogen. In a
second embodiment
Rz3 is hydrogen. In a third embodiment Rz' is hydrogen. In a further
embodiment Rz' is halo-
gen, in particular fluorine. In a still further embodiment Rz5 is hydrogen. In
a further embodi-
ment Rzfi is hydrogen. In the compound of the above formula I J is preferably
phenyl, 4-fiuoro-
10 phenyl or 2-thienyl.
fn a still further embodiment of the compound of formula I a is 1.
In a further embodiment of the compound of formula I b is 1.
In a still further embodiment of the compound of formula I c is 0.
In a further embodiment of the compound of formula I d is 0.
In a stilt further embodiment of the compound of formula I M is aryiene or
-CRz'=CRz°-, wherein Rz' and Rz8 independently from each other are
hydrogen or C,.6-alkyl,
optionally substituted with aryl or hetaryl. In one embodiment M is arylene,
in particular
phenylene. In another embodiment M is -CRz'=CRze-, wherein Rz' and Rze are
independently
hydrogen or C,~-alkyl. In a further embodiment Rz' is hydrogen. In a still
further embodiment
Rz' is C,~-alkyl, in particular methyl. In a further embodiment Rz° is
hydrogen. 1n a further
embodiment M is the E-isomer of -CRz'=CRze-. In the compound of the above
formula I M is
preferably ethenyfene, 1,3-phenylene or 1,2 -propenyVene.
In a further embodiment of the compound of formula 1 a is 0.
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In a still further embodiment of the compound of formula I a is 1.
In a further embodiment of the compound of formula I f is 0.
t In a still further embodiment of the compound of formula 1 f is 1.
PCTlDK99/00368
In a further embodiment of the compound of formula I R6 and R' are
independently from
each other hydrogen or C,.~-alkyl. 1n one embodiment R6 is hydrogen. In a
second
embodiment R6 is C,.~-alkyl, in particular methyl. In a third embodiment R' is
hydrogen. In a
further embodiment R' is C,.s-alkyl, in particular methyl.
In a still further embodiment of the compound of formula I R6 and R' or R6 and
R° or R' and R8
may optionally form -(CHZ); U-(CHZ); , wherein i and j independently from each
other are 1 or
2 and U is -O-, -S-, or a valence bond.
In a further embodiment of the compound of formula I R6 and R' form -(CHZ)~ U-
(CH2); ,
wherein i and j independently from each other are 1, 2 or 3 and U is -O-, -S-,
or a valence
bond. In one embodiment the sum i + j is 3. In a second embodiment U is a
valence bond. In
a particular embodiment (CR6R') is cyclobutyl.
in a still further embodiment of the compound of formula I R6 and R' form -
(CH2); U-(CH2); ,
wherein i and j independently from each other are 1 or 2 and U is -O-, -S-, or
a valence
bond. In one embodiment the sum i + j is 3. In a second embodiment U is a
valence bond. In
a particular embodiment (CRgR') is cyclobutyl.
In a further embodiment of the compound of formula I R° is hydrogen. In
a second
embodiment R8 is C,.~-alkyl, in particular methyl.
In a special embodiment the present invention relates to a compound of the
general formula I
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H s O ( a G Rz O
aiNw s W ~(CHR )d
R (CR R )f M ~ N ~L
R' 0 (
formula l
t
wherein
R' is hydrogen or C,.~-alkyl;
RZ is C,.~-alkyl;
L is
R9 Rio
R Rio
4
~Na a , )s -N Q
R Q or
z ( ~ r
R'z Rm R~z R~~
wherein R° is hydrogen or C,~ alkyl;
pis0or1;
q, s, t, a are independently from each other 0, 7 , 2, 3 or 4;
ris0or1;
the sum q + r + s + t + a is 0, 1, 2, 3, or 4;
R9, R'°, R", and R'z are independently from each other hydrogen or C,.~
alkyl;
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Q is >N-R'3 or
R14
'~~ T .
0
wherein o is 0, 1 or 2;
t
T is -N(R'S)(R's) or hydroxyl;
R'3, R'S, and R's are independently from each other hydrogen or C,.a alkyl;
R" is hydrogen, aryl or hetaryl;
G is
R" ,~ R,e
R,a R
or
Rz, ~ R,e ~ ~ ;
Rzo
wherein R", R'e, R's, Rz° and Rz' independently from each other are
hydrogen, halogen, aryl,
hetaryl, C,.a-alkyl or C,~-alkoxy;
J is
R~ Rzz Rz3 Rzz
Rz3 / Rz3
Rzs W I Rza W y Or ;
S
R~
wherein Rzz, Rte, Rz°, Rzs and Rzs independently from each other are
hydrogen, halogen, aryl,
hetaryl, C,~-alkyl or C,~-alkoxy;
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a is 0, 1, or 2;
b is 0, 1, or 2;
c is 0, 1, or 2;
dis0or1;
a is 0, 1, 2, or 3;
fis0orl;
RS is hydrogen or C,~-alkyl optionally substituted with one or more hydroxyl,
aryl or hetaryl;
R6 and R' are independently from each other hydrogen or C,~-alkyl, optionally
substituted
with one or more halogen, amino, hydroxyl, aryl, or hetaryi;
R8 is hydrogen or C,.~-alkyl, optionally substituted with one or more halogen,
amino, hy-
droxyl, aryl, or hetaryl;
25
R6 and R' or R6 and R8 or R' and RB may optionally form -(CHZ); U-(CHZ)~ ,
wherein i and j
independently from each other are 1, 2 or 3 and U is -O-, -S-, or a valence
bond;
M is arylene or -CRZ'=CRS-;
RZ' and R~° are independently from each other hydrogen or C,.~-alkyl,
optionally substituted
with one or more aryl or hetaryl;
or a pharmaceutically acceptable salt thereof.
Preferred compounds of formula I of the invention are:
(2E)-5-Amino-5-methylhex-2-enoic acid N-((1R)-1-{N-[(1R)-1-benzyl-2-(4-
((dimethylamino)-
methyl)piperidin-1-yl)-2-oxoethylJ-N-methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-
methyiamide
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/ /
H3C CH3 O CH3 O
HZN ~ N~N N CH3
CH3 O NCH
s
(2E)-5-Amino-5-methylhex-2-enoic acid N-((1 R)-1-{N-[(1 R)-1-benzyl-2-((3S)-3-
(dimethyl-
aminomethyl)piperidin-1-yl)-2-oxoethyl}-N-methylcarbamoyl}-2-(2-
naphthyl)ethyl)-N-
methylamide
/ /
H3C CH3 O CH3 O
HZN ~ N~ N N
CH3 O
N~CH3
I
CH3
(2E)-4-(1-Aminocyclobutyl)but-2-enoic acid N-((1 R)-1-{N-[(1 R)-1-benzyl-2-
((3S)-3-(dimethy!-
aminomethyl)piperidin-1-yl)-2-oxoethyl}-N-methylcarbamoyl}-2-(2-
naphthyl)ethyl)-N-methyl-
10 amide
/ /
O CH3 O
HZN ~ N~N N
CH3 O
N~CHs
I
CH3
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(2E)-5-Amino-5-methylhex-2-enoic acid N-((1 R)-1-{N-[(1 R)-1-benzyl-2-((2S)-2-
((dimethyl-
amino)methyl)pyrrolidin-1-yl)-2-oxoethyl]-N-methylcarbamoyl}-2-(2-
naphthyl)ethyl)-N-
methylamide
/ /
O CH3 O
H3C CH3
H2N ~ N~N N
CH3 O
N~CH3
CH3
N-((1 R)-1-{N-[(1 R)-1-Benzyl-2-((2S)-2-((dimethylamino)methyl)pyrrolidin-1-
yl)-2-oxoethyl]-N-
methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-methyl-3-
((methylamino)methyl)benzamide
/ /
O CH3 O
HN ~ N~N N
CH3 I / CH3 O
N~CH3
CH3
15
(2E)-5-Amino-5-methylhex-2-enoic acid N-((1R)-1-{N-[(1R)-1-benzyl-2-(4-
(dimethyl-
amino)piperidin-1-yl)-2-oxoethyl]-N-methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-
methylamide.
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H3C CH3 O CH3 O
H2N \ N~N
I II N
CH3 O ' ~ ~CH3
N
i
CH3
(2E)-5-Amino-5-methylhex-2-enoic acid N-methyl-N-[(1R)-1-(N-methyl-N-{(1R)-1-
[N-methyl-
N-(1-methylpiperidin-4-yl)carbamoyl]-2-phenylethyl}carbamoyl)-2-(2-
naphthyl)ethyl]amide
O CH O ~CH3
H3C CH3 a ~N
H2N \ N~N
I II -N
I
CH3 O CH3
3-Aminomethyl-N-((1 R)-1-{N-[(1 R)-1-benzyl-2-(4-methylpiperazin-1-yl)-2-
oxoethyl]-N-
methylcarbamoyl}-2-(2-naphthyl}ethyl)-N-methylbenzamide
O CH3 O
H2N \ N II N N~
CH3 O \/N'CH
3
(2E)-5-Amino-5-methylhex-2-enoic acid N-{(1 R)-1-{N-[(1 R)-1-benzyl-2-(4-
methylpiperazin-1-
yl)-2-oxoethyl]-N-methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-methylamide
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H3C CH3 O CH3 O
HZN \ N II N N
CH3 O ~N~CH
3
(2E)-5-Amino-5-methylhex-2-enoic acid N-methyl-N-((1R)-1-{N-methyl-N-[(1R)-2-
phenyl-1-
((2,2,6,6-tetramethylpiperidin-4-yl)carbamoyl)ethyl]carbamoyl}-2-{2-
naphthyl)ethyl)amide
H3C CH3
H3C CH3 O CH3 O _NH
~N CHs
HZN CH IOI H CH3
3
3-Aminomethyl-N-methyl-N-((1 R)1-{N-methyl-N-[(1 R)-2-phenyl-1-((2,2,6,6-
tetramethyl-
piperidin-4-yl)carbamoyl)ethyl]carbamoyl}-2-(2-naphthyl)ethyl)benzamide
H3C
O CH3 O CH
3
HZN I ~ N~N H NH
/ CH3 O CH
3
/ ~ H3C
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(2E)-5-Amino-3,5-dimethylhex-2-enoic acid N-methyl-N-((1 R)-1-{N-methyl-N-[(1
R)-2-phenyl
1-((2,2,6,6-tetramethylpiperidin-4-yl)carbamoyl)ethyl)carbamoyl}-2-(2-
naphthyl)ethyl)amide
H3C CH CHs O CHs O HsC CHs
HZN \ N~N H NH
CHs O CHs
HsC
(2E)-4-( 1-Aminocyclobutyl)but-2-enoic acid N-(( 1 R) 1-{N-[( 1 R) 1-benzyl-2-
(4-methylpiperazin-
1-yl)-2-oxoethyl]-N-methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-methylamide
/
O CHs O
HZN \ N 11 N N
CHs O ~N~CH
3
(2E)-5-Amino-3,5-dimethylhex-2-enoic acid N-((1R)1-{N-[(1R)1-benzyl-2-(4-
methylpiperazin-
1-yl}-2-oxoethyl)-N-methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-methylamide
H3C CH3CHs O CHs O
HZN \ N Il N N
CHs O ~N~CH
3
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(2E)-4-(1-Aminocyclobutyl)but-2-enoic acid N-((1 R)-1-{N-[(1 R)-1-benzyl-2-(4-
hydroxy-
piperidin-1-yl}-2-oxoethyl~-N-methylcarbamoyl}-2-(biphenyl-4-yl)ethyl)-N-
methylamide
/ \
/ \
O CH3 O
H2N ~ N~N N
CH3 O OH
\ /
(2E)-5-Amino-5-methylhex-2-enoic acid N-((1 R)-1-{N-[(1 R)-1-benzyl-2-(4-
hydroxypiperidin-1-
yl)-2-oxoethyl)-N-methylcarbamoyl}-2-{biphenyl-4-yl)ethyl)-N-methylamide
/ \
H3C CH3 O ' CH3 O
H2N ~ N~N N
CH3 O OH
\ /
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(2E)-5-Amino-3,5-dimethylhex-2-enoic acid N-((1 R)-1-{N-((1 R)-1-benzyl-2-(4-
hydroxypiperidin-1-yl)-2-oxoethyl]-N-methylcarbamoyl)-2-(biphenyl-4-yl)ethyl)-
N-
methylamide
H3C CHCHs O CH3 O
H2N \ N ~ N N
CH3 O OH
(2E)-5-Amino-5-methylhex-2-enoic acid N-((1R)-1-{N-((1R)-1-benzyl-2-(4-
hydroxypiperidin-1-
yl)-2-oxoethyl}-N-methylcarbamoyl}-2-(2-naphthyi)ethyl)-N-methylamide
H3C CH3 O CH3 O
H2N ~ N~N N
i
CH3 O OH
15
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(2E)-5-Amino-3,5-dimethylhex-2-enoic acid N-((1 R)-1-~N-[(1 R)-1-benzyl-2-(4-
hydroxy-
piperidin-1-yl)-2-oxoethyl]-N-methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-
methylamide
H C CH CHs O CH3 O
3 3
H2N \ N~N N
CH3 O
OH
(2E)-4-(1-Aminocyclobutyl)but-2-enoic acid N-((1 R)-1-{N-[(1 R)-1-benzyl-2-(4-
hydroxy-
piperidin-1-yl)-2-oxoethyfj-N-methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-
methylamide
O CH3 O
HZN ~ N ~ N N
CH3 O
OH
(2E)-5-Amino-5-methylhex-2-enoic acid N-((1R}-1-{N-[(1R)-1-(4-fluorobenzyl)-2-
(4-hydroxy-
piperidin-1-yf)-2-oxoethylj-N-methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-
methylamide
/ /
H3C CH3 O CH3 O
H2N ~ N~N N
CH3 O OH
F
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(2E)-5-Amino-3,5-dimethylhex-2-enoic acid N-((1R)-1-{N-[{1R)-1-(4-
fluorobenzyl)-2-(4-
hydroxypiperidin-1-yl)-2-oxoethyl]-N-methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-
methylamide
H3C CH CH3 O CH3 O
H2N ~ N~N N
CH3 O OH
F
(2E)-5-Amino-5-methylhex-2-enoic acid N-((1 R)-1-{N-[(1 R}-1-benzyl-2-(4-
hydroxy-4-(2-
thienyl)piperidin-1-yl)-2-oxoethyl]-N-methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-
methylamide
H C CH O CH3 O
3 3
HzN ~ N II N N
CH3 O S
OH
{2E)-5-Amino-5-methylhex-2-enoic acid N-{(1 R)-1-{N-[(1 R)-1-(3-
hydroxycyclohexyl-
carbamoyl)-2-phenylethyl]-N-methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-
methylamide
H3C CH3 O CH3 O
H2N ~ N~N H OH
CH3 O
1~
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(2E)4-(1-Aminocyclobutyl)but-2-enoic acid N-((1R)-1-{N-[(1R)-1-benzyl-2-(4-
(dimethyl-
amino)piperidin-1-yl)-2-oxoethylj-N-methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-
methylamide
/ \
O CH3 O
H2N ~ N N
N
CH3 O ' ~ ~CH3
N
CH3
(2E)-5-Amino-5-methylhex-2-enoic acid N-((1R)-1-{N-[(2R)-2-(4-hydfoxypiperidin-
1-yi)-2-oxo-
1-((2-thienyl)methyl)ethylj-N-methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-
methylamide
H3C CH3 O CH3 O OH
H2N \ N I I N N J
CH3 O
S
(2E)-5-Amino-3,5-dimethylhex-2-enoic acid N-((1R)-1-{N-[(2R)-2-(4-
hydroxypiperidin-1-yl)-2-
oxo-1-((2-thienyl)methyl)ethylj-N-methylcarbamoyi}-2-(2-naphthyl)ethyl)-N-
methylamide
H C CH3CH3 O CH3 O OH
3
H2N \ N II N N
CH3 O
S
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(2E)-5-Amino-5-methylhex-2-enoic acid N-((1 R)-2-(biphenyl-4-yl)-1-{N-[(2R)-2-
(4-hydroxy-
piperidin-1-yl)-2-oxo-1-((2-thienyl)methyl)ethylJ-N-methylcarbamoyl}ethyl)-N-
methylamide
H C CH O ~ CH3 O OH
3 3
H2N ~ N ~ N
CH3 O
iJ
S
(2E)-5-Amino-3,5-dimethylhex-2-enoic acid N-((1 R)-2-(biphenyl-4-yl)-1-{N-[(1
R)-2-(4-
hydroxypiperidin-1-yl)-2-oxo-1-((2-thienyl) methyl)ethyl]-N-
methylcarbamoyl}ethyl)-N-
methylamide
H3C CHCH3 O l CH3 O OH
HzN \ N~N N
CH3 O
i
S
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(2E)-5-Methyl-5-(methyiamino)hex-2-enoic acid N-((1R)-1-{N-[(1R)-1-benzyf-2-(4-
hydroxypiperidin-1-yl)-2-oxoethyl?-N-methyicarbamoyl}-2-(biphenyl-4-yl)ethyl}-
N-
methylamide
i
i
CH3 ~ CHs O
N
HN CH3 N~ N OH
CH3 CH3 O
(2E)-4-(1-Aminocyclobutyl)but-2-enoic acid ((1R)-1-{N-[(1R}-1-benzyl-2-(4-
hydroxypiperidin-
1-yl)-2-oxoethyl)-N-methylcarbamoyl}-2-(biphenyl-4-yl)ethyl)amide
/ \
/ \
O CH3 O
H2N ~ H II N N
O OH
\ /
and pharmaceutically acceptable salts thereof.
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General Methods
The methods illustrated in below schemes I - III are by no mean intended to
limit the present
invention in any aspect, but should only be seen as a guidance for how the
present com-
pounds may be prepared.
Scheme I
R9 R'° R9 ,o
R
~ ~r
' q ' reduction
BOC-N O esterification BOC-N O
v
OH
R,2 R" R,2 R" CH3
09 ,n
R R, o
oxidation a ~ rr ' reductive amination
BOC- BOC-N H
~H
o
n r< R,Z R"
R~ R'° Rs R,o
r
4 ' r
BOC-N deprotection ° '
HN
,s
~N-R
R~2 R" R's C t N-R,s
R~2 R" R's
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Amines of type
R\ R'o
(y~t N_R,s
R~2 R" R~5
may be synthesized from a BOC-protected amino acid (cf. scheme I). The acid is
trans-
formed into an ester by reaction with or without a reagent such as e. g. 1-
hydroxybenzo-
triazole, 1-hydroxy-7-azabenzotriazole, or 3-hydroxy-1,2,3-benzotriazole-4(3H)-
one and a
reagent such e.g. as N-(3-dimethyiaminopropyl)-N~-ethylcarbodiimide
hydrochloride or diiso-
propylcarbodiimide and a catalyst such as N,N-dimethylaminopyridine. The ester
may be re-
duced by a suitable reagent, such as diisobutylaluminum hydride in a
appropriate solvent
such as e.g. toluene, dichloromethane, ether, or tetrahydrofuran to give the
BOC-protected
aldehyde or alcohol. If the alcohol is obtained, the alcohol may be oxidized
to the corre-
sponding aldehyde, by a suitable method, such as e.g. dimethylsulfoxideloxalyl
chloride/ tri-
ethylamine or dimethylsufloxidelsulfotrioxidelpyridine, pyridinium dichromate,
or pyridinium
chiorochromate. A reductive amination with an appropriate amine N(R'S)(R's)
and a suitable
reagent, such as sodium cyanoborohydride or sodium triacetoxyborohydride in a
suitable
solvent such as e.g. alcohols may yield the BOC-protected amine. If at least
one of R'S or
R's is hydrogen, the amino group may be protected by a method known to those
skilled in
the art and described in the literature as e.g. T. W. Greene, P. G. M. Wuts
Protective groups
in organic synthesis, 2"° edition, Wiley, New York, before carrying out
the following steps.
The removal of the BOC-protection group can be achieved by a method known to
those
skilled in the art as described in T. W. Greene, P. G. M. Wuts Protective
groups in organic
synthesis, 2"° edition, Wiley, New York, such as e.g. hydrogen chloride
in ethyl acetate, or
trifluoroacetic acid in dichloromethane.
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Scheme II
RZ O
protection group-.N
'OH
R R,o 1~) Rz O R9 R,o
( J
HN ~q ' ~)s coupling conditions HN N
Ra a 4 Ro Qs
2.) deprotection ( J r
R'2 R" R,2~
R
1.) G
(a
protection group~N OH
G
O ( a RZ O R R,o
coupling conditions HN N N ~q '~)s
R' O ( R4 Q
2.) deprotection J ,
R'2 R"
Protection group O
..1.) RB~N~(CR6R~)~M~(CHRS)d~OH
coupling conditions
2.) deprotection
H 5 O ( aG R2 O R9 R,o
Re~N~(CRsR~)~M~(CHR.)d~N N N q )s
R' O ( J R' Q
r
R,z R"
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Compounds of the type of formula I may be synthesized by coupling of an amine
of type
Rs
~~o
Ra
~N
H
r
R' '
and a suitable protected acid with or without a coupling reagent such as e.g.
1-hydroxy-
5 benzotriazole, 1-hydroxy-7-azabenzotriazole, or 3-hydroxy-1,2,3-
benzotriazole-4(3H)-one
and a reagent such e.g. as N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide
hydrochloride or
diisopropylcarbodiimide in a suitable solvent, such as N,N-dimethylformamide
or dichloro-
methane (cf. scheme ll). The product may be deprotected at the nitrogen of the
acid by a
method known for a person skilled in the art and described in the literature
e.g. in T. W.
10 Greene, P. G. M. Wuts Protective groups in organic synthesis, 2"d edition,
Wiley, New York.
The product is coupled with a suitable protected acid with or without a
coupling reagent such
as e.g. 1-hydroxybenzotriazole, 1-hydroxy-7-azabenzotriazole, or 3-hydroxy-
1,2,3-benzo-
triazole-4(3H)-one and a reagent such e.g. as N-(3-dimethylaminopropyl)-N'-
ethyl-
carbodiimide hydrochloride or diisopropylcarbodiimide in a suitable solvent,
such as N,N-
15 dimethylformamide or dichloromethane. The product may be deprotected at the
nitrogen of
the acid by a method known for a person skilled in the art and described in
the literature e.g.
in T. W. Greene, P. G. M. Wuts Protective groups in organic synthesis,
2"° edition, Wiley,
New York. The product is coupled with suitable protected acid with or without
a coupling
reagent such as e.g. 1-hydroxybenzotriazole, 1-hydroxy-7-azabenzotriazole, or
3-hydroxy-
20 1,2,3-benzotriazole-4(3H)-one and a reagent such e.g. as N-(3-
dimethylaminopropyl)-N'-
ethylcarbodiimide hydrochloride or diisopropylcarbodiimide in a suitable
solvent, such as
N,N-dimethylformamide or dichloromethane. All protection groups may be removed
by a
method known for a person skilled in the art and described in the literature
e.g. in T. W.
Greene, P. G. M. Wuts Protective groups in organic synthesis, 2"°
edition, Wiley, New York.
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Scheme III
R2 O
R9 Rya protection group--N OH R9 Rio
1.) z
( ~ R O q v ir~s
HN coupling conditions HN N Q
Q
2.) deprotection ( ~ '
R~z R" R,z R"
1.) G
/a
protection group~N OH R9 R,o
G
I a Rz O q r s
R' O
coupling conditions HN N N Q
R' O
2.) deprotection ( J '
R~z R"
' Protection group O
..1.) Re~N~(CRsR~)~M~(CHRS)d~OH
coupling conditions
2.) deprotection
O a R' O q r s
RB~N~(CR6R7)~11A~(CHRS)d~N N N
R O ( ~ c
R~z ~R"
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Compounds of the type of formula I may be synthesized by coupling of an amine
of type
Rio
4 s
HN Q
c
and a suitable protected acid with or without a coupling reagent such as e.g.
1-hydroxy
benzotriazole, 1-hydroxy-7-azabenzotriazole, or 3-hydroxy-1,2,3-benzotriazole-
4(3H)-one
and a reagent such e.g. as N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide
hydrochloride or
diisopropylcarbodiimide in a suitable solvent, such as N,N-dimethylformamide
or dichloro-
methane (cf. scheme III). The product may be deprotected at the nitrogen of
the acid by a
method known for a person skilled in the art and described in the literature
e.g. in T. W.
Greene, P. G. M. Wuts Protective groups in organic synthesis, 2"°
edition, Wley, New York.
The product is coupled with a suitable protected acid with or without a
coupling reagent such
as e.g. 1-hydroxybenzotriazole, 1-hydroxy-7-azabenzotriazole, or 3-hydroxy-
1,2,3-benzo-
triazole-4(3H)-one and a reagent such e.g. as N-(3-dimethylaminopropyl)-N'-
ethyl-
carbodiimide hydrochloride or diisopropylcarbodiimide in a suitable solvent,
such as N,N-
dimethylformamide or dichloromethane. The product may be deprotected at the
nitrogen of
the acid by a method known for a person skilled in the art and described in
the literature e.g.
in T. W. Greene, P. G. M. Wuts Protective groups in organic synthesis,
2"° edition, Wiley,
New York. The product is coupled with suitable protected acid with or without
a coupling
reagent such as e.g. 1-hydroxybenzotriazole, 1-hydroxy-7-azabenzotriazole, or
3-hydroxy-
1,2,3-benzotriazole-4(3H}-one and a reagent such e.g. as N-(3-
dimethylaminopropyl)-N'-
ethylcarbodiimide hydrochloride or diisopropylcarbodiimide in a suitable
solvent, such as
N,N-dimethylformamide or dichloromethane. All protection groups may be removed
by a
method known for a person skilled in the art and described in the literature
e.g. in T. W.
Greene, P. G. M. Wuts Protective groups in organic synthesis, 2"°
edition, Wiley, New York.
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The compounds of formula 1 exhibit an improved resistance to proteolytic
degradation by
enzymes because they are non-natural, in particular because the natural amide
bonds are
replaced by non-natural amide bond mimetics. The increased resistance to
proteolytic
degradation of the compounds of the invention in comparison with known.
hormone releasing
peptides is expected to improve their bioavailability compared to that of the
peptides suggested
in the prior literature.
In the above structural formulas and throughout the present specification, the
following terms
have the indicated meanings:
The C,.~-alkyl, C,.~-alkylene, C,.,-alkyl or C,.~-alkylene groups specified
above are intended to
include those alkyl or alkyiene groups of the designated length in either a
linear or branched or
cyclic configuration. Examples of linear alkyl are methyl, ethyl, propyl,
butyl, pentyl, and hexyl
and their corresponding divalent moieties, such as ethylene. Examples of
branched alkyl are
isopropyl, sec-butyl, tent-butyl, isopentyl, and isohexyl and their
corresponding divalent moie-
ties, such as isopropylene. Examples of cyclic alkyl are C~-cycloalkyl such as
cyclopropyl, cy-
clobutyl, cyGopentyl and cyclohexyl and their corresponding divalent moieties,
such as cyclo-
propylene.
The C,.~-alkoxy groups specified above are intended to include those alkoxy
groups of the
designated length in either a linear or branched or cyclic configuration.
Examples of linear alk-
oxy are methoxy, ethoxy, propoxy, butoxy, pentoxy, and hexoxy. Examples of
branched alkoxy
are isopropoxy, sec-butoxy, tent-butoxy, isopentoxy, and isohexoxy. Examples
of cyclic alkoxy
are C~-cycloalkoxy such as cyclopropyloxy, cyclobutyloxy, cyclopentyloxy and
cyclohexyloxy.
In the present context, the term "aryl" is intended to include monovalent
carbocyclic aromatic
ring moieties, being either monocyclic, bicyclic or poiycyclic, e.g. selected
from the group con-
sisting of phenyl and naphthyl, optionally substituted with one or more C,.~-
alkyl, C,.~-alkoxy,
halogen, amino or aryl.
In the present context, the term "arylene" is intended to include divalent
carbocyclic aromatic
ring moieties, being either monocycfic, bicyclic or poiycyclic, e.g. selected
from the group con-
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sisting of phenylene and naphthylene, optionally substituted with one or more
C,.~-alkyl, C,~-
alkoxy, halogen, amino or aryl.
In the present context, the term "hetaryl" is intended to include monovalent
heterocyclic aro-
matic ring moieties, being either monocyclic, bicyclic or polycyclic, e.g.
selected from the group
consisting of pyridyl, 1-H-tetrazol-5-yl, thiazolyl, imidazolyl, indolyl,
pyrimidinyl, thiadiazolyl,
pyrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thienyl, quinolinyl, pyrazinyl,
or isothiazolyl, option-
ally substituted with one or more C,.6-alkyl, C,.~-alkoxy, halogen, amino or
aryl.
In the present context, the term "hetarylene" is intended to include divalent
heterocyclic aro-
matic ring moieties, being either monocyclic, bicyclic or polycyclic, e.g.
selected from the group
consisting of pyridinediyl, 1-H-tetrazolediyl, thiazoldiyl, imidazolediyl,
indolediyl, pyrimidinediyl,
thiadiazolediyl, pyrazolediyf, oxazolediyl, isoxazolediyl, oxadiazolediyl,
thiophenediyl, quinolin-
ediyl, pyrazinediyl, or isothiazolediyl, optionally substituted with one or
more C,~-alkyl, C,.~-
alkoxy, halogen, amino or aryl.
In the present context, the term "heterocyclic system" is intended to include
aromatic as well as
non-aromatic ring moieties, which may be monocyclic, bicyclic or poiycyclic,
and contain in
their ring structure at least one, such as one, two or three, nitrogen
atom(s), and optionally one
or more, such as one or two, other hetero atoms, e.g. sulpher or oxygen atoms.
The hetero-
cyclic system is preferably selected from pyrazole, pyridazine, triazine,
indazole, phthalazine,
cinnoline, pyrazolidine, pyrazoline, aziridine, dithiazine, pyn-ol, imidazol,
pyrazole, isoindole,
indole, indazole, purine, pyrrolidine, pyrroline, imidazolidine, imidazoline,
pyrazolidine,
pyrazoline, piperidine, piperazine, indoline, isoindoline, or morpholine,
optionally substituted
with one or more C,~-alkyl, C,~-alkoxy, halogen, amino, oxy or aryl.
The term "halogen" is intended to include chlorine (CI), fluorine (F), bromine
(Br) and iodine (I).
In the context of the present application, the term "growth hormone
secretagogue" is intended
to include any compound which has the capacity, directly or indirectly, of
inducing (i.e. stimu-
lating or increasing) the release of growth hormone from the pituitary gland.
The term "growth
hormone secretagogue" includes growth hormone releasing peptides, growth
hormone releas-
ing peptidomimetics, and growth hormone releasing compounds of a nonpeptidyl
nature.
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The compounds of the present invention may optionally be on a pharmaceutically
acceptable
salt form such as the pharmaceutically acceptable acid addition salts of
compounds of formula
I which include those prepared by reacting the compound of formula I with an
inorganic or or-
5 ganic acid such as hydrochloric, hydrobromic, sulfuric, acetic, phosphoric,
lactic, malic, malefic,
mandelic phthalic, citric, glutaric, gluconic, methanesutfonic, salicylic,
succinic, tartaric, tolue-
nesulfonic, trifluoracetic, sulfamic or fumaric acid andlor water.
The compounds of formula I may be administered in pharmaceutically acceptable
acid addition
10 salt form or, where appropriate, as a alkali metal or alkaline earth metal
or lower alkylammo-
nium salt. Such salt forms are believed to exhibit approximately the same
order of activity as
the free base forms.
In another aspect, the present invention relates to a pharmaceutical
composition comprising,
15 as an active ingredient, a compound of the general formula I or a
pharmaceutically acceptable
salt thereof together with a pharmaceutically acceptable carrier or diluent.
Pharmaceutical compositions containing a compound of the present invention may
be pre-
pared by conventional techniques, e.g. as described in Remington's
Pharmaceutical Sciences,
20 1985 or in Remington: The Science and Practice of Pharmacy, 19th Edition
{1995). The com-
positions may appear in conventional forms, for example capsules, tablets,
aerosols, solutions,
suspensions or topical applications.
The pharmaceutical carrier or diluent employed may be a conventional solid or
liquid carrier.
25 Examples of solid carriers are lactose, terra alba, sucrose, cyclodextrin,
talc, gelatin, agar, pec-
tin, acacia, magnesium stearate, stearic acid or lower alkyl ethers of
cellulose. Examples of
liquid carriers are syrup, peanut oil, olive oil, phospholipids, fatty acids,
fatty acid amines,
polyoxyethylene or water.
30 Similarly, the carrier or diluent may include any sustained release
material known in the art,
such as glyceryl monostearate or giyceryl distearate, alone or mixed with a
wax.
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If a solid carrier is used for oral administration, the preparation may be
tabletted, placed in a
hard gelatin capsule in powder or pellet form or it can be in the form of a
troche or lozenge.
The amount of solid carrier will vary widely but will usually be from about 25
mg to about 1 g. If
a liquid carrier is used, the preparation may be in the form of a syrup,
emulsion, soft gelatin
capsule or sterile injectable liquid such as an aqueous or non-aqueous liquid
suspension or
b
solution.
A typical tablet which may be prepared by conventional tabletting techniques
may contain:
Core:
Active compound (as free compound or salt thereof) 1 Omg
Colloidal silicon dioxide 1.5mg
(Aerosil)
Cellulose, microcryst. (Avicel)70mg
Modified cellulose gum (Ac-Di-Soi)7.5mg
Magnesium stearate
Coating:
HPMC approx. 9mg
*Mywacett 9-40 T approx. 0.9mg
*Acylated monogiyceride used as plasticizer for film coating.
For nasal administration, the preparation may contain a compound of formula I
dissolved or
suspended in a liquid carrier, in particular an aqueous carrier, for aerosol
application. The car-
rier may contain additives such as solubilizing agents, e.g. propylene glycol,
surfactants, ab-
sorption enhancers such as lecithin (phosphatidylcholine) or cyclodextrin, or
preservatives such
as parabenes.
it has been demonstrated that compounds of the general formula I possess the
ability to re-
lease endogenous growth hormone in vivo. The compounds may therefore be used
in the
treatment of conditions which require increased plasma growth hormone levels
such as in
growth hormone deficient humans or in elderly patients or livestock.
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Thus, in a particular aspect, the present invention relates to a
pharmaceutical composition for
stimulating the release of growth hormone from the pituitary, the composition
comprising, as an
active ingredient, a compound of the general formula I or a pharmaceutically
acceptable salt
thereof together with a pharmaceutically acceptable carrier or diluent.
In a further aspect, the present invention relates to a method of stimulating
the release of
growth hormone from the pituitary, the method comprising administering to a
subject in need
thereof an effective amount of a compound of the general formula I or a
pharmaceutically ac-
ceptable salt thereof.
In a still further aspect, the present invention relates to a method of
treating growth retardation
in connection with asthma, the method comprising administering to a subject in
need thereof
an effective amount of a growth hormone secretagogue or a pharmaceutically
acceptable salt
thereof. In a particular embodiment, the present invention relates to a method
of treating
growth retardation in connection with asthma, the method comprising
administering to a sub-
jest in need thereof an effective amount of a compound of the general formula
I or a pharma-
ceutically acceptable salt thereof.
In a still further aspect, the present invention relates to a method of
treating growth retardation
in connection with juvenile rheumatic arthritis or systic fibrosis, the method
comprising admin-
istering to a subject in need thereof an effective amount of a growth hormone
secretagogue or
a pharmaceutically acceptable salt thereof. In one embodiment, the present
invention relates to
a method of treating growth retardation in connection with juvenile rheumatic
arthritis, the
method comprising administering to a subject in need thereof an effective
amount of a growth
hormone secretagogue or a pharmaceutically acceptable salt thereof. In a
second embodi-
ment, the present invention relates to a method of treating growth retardation
in connection
with systic fibrosis, the method comprising administering to a subject in need
thereof an effec-
tive amount of a growth hormone secretagogue or a pharmaceutically acceptable
salt thereof.
In a particular embodiment, the present invention relates to a method of
treating growth retar-
dation in connection with juvenile rheumatic arthritis, the method comprising
administering to a
subject in need thereof an effective amount of a compound of the general
formula I or a phar-
maceutically acceptable salt thereof. In another particular embodiment, the
present invention
relates to a method of treating growth retardation in connection with systic
fibrosis, the method
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38
comprising administering to a subject in need thereof an effective amount of a
compound of
the general formula I or a pharmaceutically acceptable salt thereof.
In a still further aspect, the present invention relates to the use of a
compound of the general
formula 1 or a pharmaceutically acceptable salt thereof for the preparation of
a medicament for
stimulating the release of growth hormone from the pituitary.
To those skilled in the art, it is well known that the current and potential
uses of growth hor-
mone in humans are varied and multitudinous. Thus, compounds of formula I can
be adminis-
tered for purposes stimulating release of growth hormone from the pituitary
and would then
have similar effects or uses as growth hormone itself. Compounds of formula t
are useful for:
stimulation of growth hormone release in the elderly, prevention of catabolic
side effects of glu-
cocorticoids, prevention and treatment of osteoporosis, treatment of chronic
fatigue syndrom
(CFS), treatment of acute fatigue syndrom and muscle loss following elective
surgery, stimula-
tion of the immune system, acceleration of wound healing, accelerating bone
fracture repair,
accelerating complicated fractures, e.g. disctraction osteogenesis, treatment
of wasting secon-
dary to fractures, treatment of growth retardation, treating growth
retardation resulting from re-
nal failure or insufficiency, treatment of cardiomyopathy, treatment of
wasting in connection
with chronic liver disease, treatment of thrombocytopenia, treatment of growth
retardation in
connection with Crohn's disease, treatment of short bowel syndrome, treatment
of wasting in
connection with chronic obstructive pulmonary disease (COPD), treatment of
complications
associated with transplantation, treatment of physiological short stature
inGuding growth hor-
mone deficient children and short stature associated with chronic illness,
treatment of obesity
and growth retardation associated with obesity, treatment of anorexia,
treatment of growth re-
tardation associated with the Prader Wilfi syndrome and Turner's syndrome;
increasing the
growth rate of a patient having partial growth hormone insensitive syndrome,
accelerating the
recovery and reducing hospitalization of bum patients; treatment of
intrauterine growth retarda-
tion, skeletal dysplasia, hypercortisolism and Cushing's syndrome; induction
of pulsatile growth
hormone release; replacement of growth hom~one in stressed patients, treatment
of osteo-
chondrodysplasias, Noonan's syndrome, schizophrenia, depressions, Alzheimer's
disease,
delayed wound heating and psychosocial deprivation, treatment of catabolism in
connection
with pulmonary dysfunction and ventilator dependency; treatment of cardiac
failure or related
vascular dysfunction, treatment of impaired cardiac function, treatment or
prevention of myo-
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39
cardial infarction, lowering blood pressure, protection against ventricular
dysfunction or preven-
tion of reperfusion events; treatment of adults in chronic dialysis;
attenuation of protein cata-
bolic responses after major surgery, reducing cachexia and protein loss due to
chronic illness
such as cancer or AIDS; treatment of hyperinsulinemia including
nesidioblastosis, adjuvant
treatment for ovulation induction; stimulation of thymic development and
prevention of the
age-related decline of thymic function, treatment of immunosuppressed
patients; treatments of
sarcopenia, treatment of wasting in connection with AIDS; improvement in
muscle strength,
mobility, maintenance of skin thickness, treatment of metabolic homeostasis
and renal homeo-
stasis in the frail elderly, stimulation of osteoblasts, bone remodelling and
cartilage growth;
regulation of food intake; stimulation of the immune system in companion
animals and treat-
ment of disorder of aging in companion animals, promoting growth in livestock
and stimulation
of wool growth in sheep, increasing milk production in livestock, treatment of
metabolic syn-
drom (syndrome X), treatment of insulin resistance, including NIDDM, in
mammals, e.g. hu-
mans, treatment of insulin resistance in the heart, improvement of sleep
quality and con-ection
of the relative hyposomatotropism of senescence due to high increase in REM
sleep and a de-
crease in REM latency, treatment of hypothermia, treatment of frailty
associated with aging,
treatment of congestive heart failure, treatment of hip fractures, treatment
of immune defi-
ciency in individuals with a depressed T4lT8 cell ratio, treatment of muscular
atrophy, treat-
ment of musculoskeletal impairment in elderly, enhancing the activity of
protein kinase B
(PKB), improvement of the overall pulmonary function, and treatment of sleep
disorders.
Within the context of the present application, the term "treatment" is also
intended to include
prophylactic treatment.
In a further aspect the present invention relates to the use of a growth
hormone secretagogue
or a pharmaceutically acceptable salt thereof for the preparation of a
medicament for the
treatment of growth retardation in connection with asthma. In one particular
embodiment the
invention relates to the use of a compound of the general formula I or a
pharmaceutically
acceptable salt thereof for the treatment of growth retardation in connection
with asthma. In a
second particular embodiment the invention relates to the use of growth
hormone releasing
peptides, growth hormone releasing peptidomimetics, or growth hormone
releasing
compounds of a nonpeptidyl nature or a pharmaceutically acceptable salt
thereof for the
treatment of growth retardation in connection with asthma.
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In a still further aspect the present invention relates to the use of a growth
hormone
secretagogue or a pharmaceutically acceptable salt thereof for the preparation
of a
medicament for the treatment of growth retardation in connection with juvenile
fieumatic
5 arthritis or systic fibrosis. In one embodiment the invention relates to the
use of a growth
hormone secretagogue or a pharmaceutically acceptable salt thereof for the
preparation of a
medicament for the treatment of growth retardation in connection with juvenile
rheumatic
arthritis. In a second embodiment the invention relates to the use of a growth
hormone
secretagogue or a pharmaceutically acceptable salt thereof for the preparation
of a
10 medicament for the treatment of growth retardation in connection with
systic fibrosis. In one
particular embodiment the invention relates to the use of a compound of the
general formula I
or a pharmaceutically acceptable salt thereof for the treatment of growth
retardation in
connection with juvenile rheumatic arthritis. In another particular embodiment
the invention
relates to the use of a compound of the general formula I or a
pharmaceutically acceptable salt
15 thereof for the treatment of growth retardation in connection with systic
fibrosis. In a further
particular embodiment the invention relates to the use of growth hormone
releasing peptides,
growth hormone releasing peptidomimetics, or growth hormone releasing
compounds of a
nonpeptidyl nature or a pharmaceutically acceptable salt thereof for the
treatment of growth
retardation in connection with juvenile rheumatic arthritis. In a still
further particular
20 embodiment the invention relates to the use of growth hormone releasing
peptides, growth
hormone releasing peptidomimetics, or growth hormone releasing compounds of a
nonpeptidyl
nature or a pharmaceutically acceptable salt thereof for the treatment of
growth retardation in
connection with systic fibrosis.
25 For the above indications the dosage will vary depending on the growth
hormone secretagogue
employed, e.g. on the compound of formula I employed, on the mode of
administration and on
the therapy desired. However, generally dosage levels between 0.0001 and 100
mg/kg body
weight daily are administered to patients and animals to obtain effective
release of endogenous
growth hormone. Morever the compounds of formula I have no or substantially no
side-effects,
30 when administered in the above dosage levels, such side-effects being e.g.
release of LH,
FSH, TSH, ACTH, vasopressin, oxytocin, cortisol and/or prolactin. Usually,
dosage forms suit-
able for oral, nasal, pulmonal or transdermai administration comprise from
about 0.0001 mg to
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41
about 100 mg, preferably from about 0.001 mg to about 50 mg of the compounds
of formula I
admixed with a pharmaceutically acceptable carrier or diluent.
The dosage of the compounds according to this invention is suitably 0.01-500
mg/day, e.g.
from about 5 to about 50 mg, such as about 10 mg per dose, when administered
to patients,
e.g. humans, as a drug.
Optionally, the pharmaceutical composition of the invention may comprise a
compound of for-
mula 1 combined with one or more compounds exhibiting a different activity,
e.g., an antibiotic
or other pharmacologically active material.
The route of administration may be any route which effectively transports the
active compound
to the appropriate or desired site of action, such as oral, nasal, pulmonary,
transdermal or par-
enteral, the oral route being preferred.
Apart from the pharmaceutical use of the compounds of formula I, they may be
useful in vitro
tools for investigating the regulation of growth hormone release.
Compounds of formula I may also be useful in vivo tools for evaluating the
growth hormone
releasing capability of the pituitary. For example, senrm samples taken before
and after ad-
ministration of these compounds to humans can be assayed for growth hormone.
Comparison
of the growth hormone in each serum sample would directly determine the
ability of the pa-
tients pituitary to release growth hormone.
Compounds of formula I may be administered to commercially important animals
to increase
their rate and extent of growth, and to increase milk and wool production.
A further use of growth hormone secretagogue compounds of formula I is in
combination with
other secretagogues such as GHRP (2 or 6), GHRH and its analogues, growth
hormone and
its analogues or somatomedins including IGF-1 and IGF-2.
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Pharmacological Methods
Compounds of formula I may be evaluated in vitro for their efficacy and
potency to release
growth hormone in rat pituitary primary cultures, and such evaluation may be
performed as de-
scribed below.
The isolation of rat pituitary cells is a modification of O. Sartor et al.,
Endocrinol~~ 11~, 1985,
pp. 952-957. Male albino Sprague-Dawley rats (250 +/- 25 grams) were purchased
from
Mollegaard, Lille Skensved, Denmark. The rats were housed in group cages (four
ani-
mals/cage) and placed in rooms with 12 hour light cycle. The room temperature
varied from
19-24°C and the humidity from 30 - 60%.
The rats were decapitated and the pituitaries dissected. The neurointermediate
lobes were re-
moved and the remaining tissue was immediately placed in icecold isolation
buffer (Gey's me-
~ 5 dium (Gibco 041-04030) supplemented with 0.25% D-glucose, 2% non-essential
amino acids
(Gibco 043-01140) and 1 % bovine serum albumine (BSA) (Sigma A-4503)). The
tissue was cut
into small pieces and transferred to isolation buffer supplemented with 3.8
mg/ml of trypsin
(Worthington #3707 TRL-3) and 330 mg/ml of DNase (Sigma D-4527). This mixture
was incu-
bated at 70 rotations/min for 35 min at 37°C in a 95/5% atmosphere of
O~/COZ. The tissue was
20 then washed three times in the above buffer. Using a standard pasteur
pipette, the tissue was
then aspirated into single cells. After dispersion, cells were filtered
through a nylon filter (160
mm) to remove undigested tissue. The cell suspension was washed 3 times with
isolation
buffer supplemented with trypsin inhibitor (0.75 mg/ml, Worthington #2829) and
finally resu-
spended in culture medium; DMEM (Gibco 041-01965) supplemented with 25 mM
HEPES
25 (Sigma H-3375), 4 mM glutamine (Gibco 043-05030H), 0.075% sodium
bicarbonate (Sigma S-
8875), 0.1% non-essential amino acid, 2.5% fetal calf serum (FCS, Gibco 011-
06290), 3%
horse serum (Gibco 034-06050), 10% fresh rat serum, 1 nM T3 (Sigma T-2752) and
40 mg/I
dexamethasone (Sigma D-4902) pH 7.3, to a density of 2 x 105 cells/ml. The
cells were seeded
into microtiter plates (Nunc, Denmark), 200 mUwell, and cultured for 3 days at
37°C and 8%
30 CO2.
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Compound testing
After culturing, the cells were washed twice with stimulation buffer (Hanks
Balanced Salt Solu-
tion (Gibco 041-04020} supplemented with 1 % BSA (Sigma A-4503), 0.25% D-
glucose (Sigma
G-5250) and 25 mM HEPES (Sigma H-3375) pH 7.3) and preincubated for 1 hour at
37°C. The
buffer was exchanged with 90 ml stimulation buffer (37°C). Ten ml test
compound solution was
added and the plates were incubated for 15 min at 37°C and 5% C02. The
medium was de-
canted and analyzed for GH content in an rGH SPA test system.
All compounds were tested in doses ranging from 10 pM to 100 mM. A dose-
response relation
was constructed using the Hill equation (Fig P, Biosoft). The efficacy
(maximal GH released,
E"",~ was expressed in % of the E""~ of GHRP-6. The potency (ECM) was
determined as the
concentration inducing half maximal stimulation of the GH release.
Compounds of formula I may be evaluated for their metabolic stability using
the procedure de-
scribed below:
Compounds is dissolved at a concentration of 1 mglml in water. 25 ml of this
solution is added
to 175 ml of the respective enzyme-solution (resulting in an enzymeaubstrate
ratio (wlw) of
approximately 1:5). The solution is left at 37°C overnight. 10 ml of
the various degradation so-
lotions is analyzed against a corresponding zero-sample using flow injection
electrospray mass
spectrometry (ESMS) with selected ion monitoring of the molecular ion. If the
signal has de-
creased more than 20% compared to the zero-sample, the remainder of the
solution is ana-
lyzed by HPLC and mass spectrometry in order to identify the extent and sites)
of degradation
precisely.
Several standard peptides (ACTH 4-10, Angiotensin 1-14 and Glucagon) have been
included
in the stability tests in order to verify the ability of the various solutions
to degrade peptides.
Standard peptides (angiotensin 1-14, ACTH 4-10 and glucagon) were purchased
from Sigma,
MO, USA)
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Enzymes (trypsin, chymotrypsin, eiastase aminopeptidase M and carboxypeptidase
Y and B)
were all purchased from Boehringer Mannheim GmbH (Mannheim, Germany)
Pancreatic enzyme mix: trypsin, chymotrypsin and elastase in 100 mM
ammoniumbicarbonate
pH 8.0 (all concentrations 0.025 mg/ml).
Carboxypeptidase mix: carboxypeptidase Y and B in 50 mM ammoniumacetate pH 4.5
(all
concentrations 0.025 mglml).
Aminopeptidase M solution: aminopeptidase M (0.025 mg/ml) in 100 mM
ammoniumbicarbon-
ate pH 8.0
Mass spectrometric analysis was performed using two different mass
spectrometers. A Sciex
API III triple quadrupole LC-MS instrument (Sciex instruments, Thomhill,
Ontario) equipped
with an electrospray ion-source and a Bio-Ion 20 time-of-Right Plasma
Desorption instrument
(Bio-lon Nordic AB, Uppsala, Sweden).
Quantification of the compounds (before and after degradation) was done on the
API III in-
strument using single ion monitoring of the molecular ion in question with
flow injection of the
anaiyte. The liquid flow (MeOH:water 1:1 ) of 100 mUmin was controlled by an
ABI 1408 HPLC
unit (Perkin-Elmer Applied Biosystems Divisions, Foster City, CA). The
instrument parameters
were set to standard operation conditions, and SIM monitoring was performed
using the most
intense molecular ion (in most cases this corresponded to the doubly charged
molecular ion).
Identification of degradation products furthermore involved the use of plasma
desorption mass
spectrometry (PDMS) with sample application on nitrocellulose coated targets
and standard
instrumental settings. The accuracy of the hereby determined masses is
generally better than
0.1 %.
Separation and isolation of degradation products was done using a HY-TACH C-18
reverse
phase 4.6x105 mm HPLC column (Hewlett-Packard Company, Palo Alto, CA) with a
standard
acetonitril: TFA separation gradient. The HPLC system used was HP1090M
(Hewlett-Packard
Company, Palo Alto, CA).
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Peptide de- MW/SIM Carboxy- Pan.
rivative ion (amu) peptidaseenzyme
mix mix
Standards
ACTH 4-10 1124.5/562.+ -
8
Glucagon 3483/871.8- -
Insulin (B23-859.1/430.6
29)
Angiotensin 1760.11881.- -
1- 0
14
GHRP-2 817.4!409.6- -
GHRP-6 872.6/437.4- -
+: Stable (less than 20% decrease in SIM signal after 24 h in degradation
solution)
-: Unstable (more than 20% decrease in SIM signal after 24 h in degradation
solution)
5
Any novel feature or combination of features described herein is considered
essential to this
invention.
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EXAMPLES:
The process for preparing compounds of formula I and preparations containing
them is fur-
ther illustrated in the following examples, which however, are not to be
construed as limiting.
The structures of the compounds are confirmed by either elemental analysis
(MA) nuclear
b
magnetic resonance (NMR) or mass spectrometry (MS). NMR shifts (d) are given
in parts
per million (ppm) and only selected peaks are given. mp is melting point and
is given in °C.
Column chromatography was carried out using the technique described by W.C.
Still et al,
J. Org. Chem. 1978, 43, 2923-2925 on silica gel 60. Compounds used as starting
materials
are either known compounds or compounds which can readily be prepared by
methods
known per se.
HPLC-Analysis:
Method A1.
The RP-analysis was performed using UV detections at 214, 254, 276, and 301 nm
on a
218TP54 4.6 mm x 250 mm 5m C-18 silica column (The Seperations Group,
Hesperia),
which was eluted at 1 mUmin at 42°C. The column was equilibrated with
5% acetonitrile in a
buffer consisting of 0.1 M ammonium sulfate, which was adjusted to pH 2.5 with
4M sulfuric
acid. after injection the sample was eluted by a gradient of 5% to 60%
acetonitrile in the
same buffer during 50 min.
Method B1.
The RP-analysis was performed using UV detections at 214, 254, 276, and 301 nm
on a
218TP54 4.6 mm x 250 mm 5m C-18 silica column (The Seperations Group,
Hesperia),
which was eluted at 1 mUmin at 42°C. The column was equilibrated with
5% (acetonitrile +
0.1 % TFA) in an aqueous solution of TFA in water (0.1 %). After injection the
sample was
eluted by a gradient of 5% to 60% (acetonitrile + 0.1 % TFA) in the same
aqueous buffer
during 50 min.
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Abbreviations:
TLC: thin layer chromatography
DMSO: dimethylsulfoxide
min: minutes
h: hours
Boc: tert butyloxycarbonyl
d
DMF: dimethylformamide
THF: tetrahydrofuran
EDAC: N-ethyl-N'-dimethylaminopropylcarbodiimide hydrochloride
HOAt:1-hydroxy-7-azabenzotriazole
DIEA: diisopropylethylamine
TFA: trifluoroacetic acid
Buildingblocks:
'
N-methylated aminoacids used in the following examples were prepared as in
Can. J. Chem.
1977, 55, 906.
3-Hydroxy-1,1-dimethylpropylcarbamic acid tert-butyl ester.
CH3 O H3C CH3
H3C~ ~ ~~~OH
H3C O H
At 0 °C, ethyl chloroformate (1.10 mL, 11.5 mmol) was given dropwise to
a solution of 3-tert-
butoxycarbonylamino-3-methylbutanoic acid (2.50 g, 11.5 mmol) and
triethylamine (1.92 mL,
13.8 mmol) in tetrahydrofuran (10 mL). The solution was stirred for 40 min at
0 °C. The formed
precipitate was filtered off and washed with tetrahydrofuran (20 mL). The
liquid was immediate-
ly cooled to 0 °C. A 2M solution of lithium boronhydride in
tetrahydrofuran (14.4 mL, 28.8
mmol) was added dropwise. The solution was stirred at 0 °C for 2 h, and
then warmed to room
temperature. over a period of 4 h. It was cooled to 0 °C. Methanol (5
mL) was added carefully.
1N Hydrochloric acid (100 mL) was added. The solution was extracted with ethyl
acetate (2 x
100 mL, 3 x 50 mL). The combined organic layers were washed with saturated
sodium
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hydrogen carbonate solution (100 mL) and dried over magnesium sulfate. The
solvent was
removed in vacuo. The crude product was chromatographed on silica (110 g) with
ethyl
acetate/heptane 1:2 to give 1.84 g of 3-hydroxy-1,1-dimethylpropylcarbamic
acid tert-butyl
ester.
'H-NMR (CDCl3): d 1.33 (s, 6 H); 1.44 (s, 9 H); 1.88 (t, 2 H); 1.94 (br, 1 H);
3.75 (q, 2 H); 4.98
(br, 1 H).
t
3-(tert-Butoxycarbonylamino)-3-methylbutanal:
CH3 O H3C CH3 H
HsC~O~N~~~
HC H O
DMSO (1.22 mL, 17.2 mmol) was added to a solution of oxalyl chloride (1.1 mL,
12.9 mmol) at
-78 °C in dichloromethane (15 mL). The mixture was stirred for 15 min
at -78 °C. A solution of
3-hydroxy-1,1-dimethylpropylcarbamic acid tert-butyl ester (1.75 g, 8.6 mmol)
in dichloro-
methane (10 mL) was added dropwise over a period of 15 min. The solution was
stirred at -78
°C for another 15 min. Triethylamine (6.0 mL, 43 mmol) was added. The
solution was stirred at
-78 °C for 5 min and then warmed to room temperature. The solution was
diluted with dichloro-
methane (100 mL) and extracted with 1 N hydrochloric acid (100 mL). The
aqueous phase was
extracted with dichloromethane (50 mL). The combined organic layers were
washed with
saturated sodium hydrogen carbonate solution (100 mL) and dried over magnesium
sulfate.
The solvent was removed in vacuo. The crude product was purfied by column
chromatography
on silica (140 g) with ethyl acetate/heptane (1:3) to give 1.10 g of 3-(tert-
butoxycarbonylamino)-
3-methylbutanal.
MHz-'H-NMR (CDCI3): d 1.39 (s, 6 H); 1.45 (s, 9 H); 2.85 (d, 2 H); 4.73 (br. 1
H); 9.80 (t, 1 H).
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Ethyl (2E)-5-{tert-Butoxycarbonylamino)-5-methylhex-2-enoate:
CH3 O H3C CH3 O
H3C-~O~N \ O~CH
H3C H
b
Triethyiphoshonoacetate (1.96 ml, 9.8 mmol) was dissolved in tetrahydrofuran
(30 ml).
Potassium tert-butoxide (1.10 g, 9.8 mmol) was added. The solution was stirred
for 40 min at
room temperature. A solution of 3-(tent-butoxycarbonylamino)-3-methylbutanal
(1.10 g, 5.5
mmol) in Tetrahydrofuran (6 m1) was added. The solution was stirred at room
temperature. for
75 min. It was diluted with ethyl acetate (100 ml) and 1 N hydrochloric acid
(100 ml). The
1 o phases were separated. The aqueous phase was extracted with ethyl acetate
{2 x 50 ml). The
combined organic phases were washed with saturated sodium hydrogen carbonate
solution
(60 ml) and dried over magnesium sulfate. The solvent was removed in vacuo.
The crude
product was purfied by column chromatography on silica (90 g) with ethyl
acetate/hepatane
(1:4) to give 1.27 g of ethyl (2E)-5-(tert-butoxycarbonylamino)-5-methylhex-2-
enoate.
'H-NMR (CDC13): d 1.30 (s, 6 H); 1.30 (t, 3 H); 1.46 (s, 9 H); 2.62 (d, 2 H);
4.27 (q, 2 H); 4.42
(br, 1 H); 5.88 (d, 1 H); 6.94 (td, 1 H).
{2E)-5-(tert-Butoxycarbonylamino)-5-methylhex-2-enoic acid:
CH3 O H3C CH3 O
H3C~O~N \ OH
H3C H
Ethyl (2E)-5-(tert-butoxycarbonylamino~5-methylhex-2-enoate (1.233 g, 4.54
mmol) was
dissolved in dioxane (20 ml). Lithium hydroxide (0.120 g, 5.00 mmol) was added
as a solid.
Water (10 ml) was added, until a clear solution was reached. The solution was
stirred 16 h at
room temperature. The solution was diluted with water (70 ml) and was
extracted with tert-butyl
methyl ether (2 x 100 ml). The aqueous phase was acidfied with 1 N sodium
hydrogensulfate
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solution (pH = 1 ) and was extracted with tert-butylmethylether (3 x 70 ml).
The organic phases
were combined and dried over magnesium sulfate. The solvent was removed in
vacuo to give
1.05 g of (2E}-5-(tert-butoxycarbonylamino}-5-methylhex-2-enoic acid. The
crude product was
used for further syntheses.
5 'H-NMR (DMSO d6): d 1.15 (s, 6 H); 1.35 (s, 9 H); 2.53 (d, 2 H); 5.75 (d, 1
H); 6.57 (br, 1 H);
6.75 (td, 1 H); 12.15 (s, 1 H).
b
1-Aza-spiro[3.3]heptan-2-one:
NH
1a O
Methylenecyclobutane (40.Og, 0.587 mol) was dissolved in diethylether (250
ml). At - 40 °C ,
chlorosulfonylisocyanate (26 ml, 0.294 mol) was added dropwise. The reaction
mixture was
warmed to 10 °C, An exothermic reaction was observed, and precipitation
was formed. The
15 reaction mixture was cooled to -20 °C. It was stirred for 16 h,
while it was warming up to
room temperature. A saturated aqueous solution of sodium sulfite (100 ml) was
added drop-
wise. The reaction mixture was stirred vigorously for 1 h. Another saturated
aqueous solution
of sodium sulfite (100 ml) was added dropwise. Solid sodium hydrogen carbonate
was
added, until pH 7. Dichloromethane (500 ml) was added. The phases were
separated. The
20 organic layer was dried over magnesium sulfate. The solvent was removed in
vacuo, to give
23.59 g of 1-aza-spiro[3.3]heptan-2-one.
'H-NMR (CDCI3): d 1.75 (m, 2 H); 2.26 (m, 2 H); 2.39 (m, 2 H); 2.96 (s, 2 H);
6.55 (br, 1 H).
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2-Oxo-1-azaspiro[3.3]heptane-1-carboxylic acid fart-butylester
N
p ~O
O CH3
~CH3
CH3
A solution of di-tart butyl Bicarbonate (55.7 g, 0.211 mol) in dichloromethane
(100 ml) was
added dropwise to a solution of 1-aza-spiro[3.3]heptan-2-one, triethylamine
(36 ml, 0.255
mot), and 4-dimethyiaminopyridiene (2.6 g, 0.021 mol) in dichloromethane (100
ml). The re-
action mixture was stirred for 16 h at room temperature. It was washed with a
10% aqueous
solution of ammonium chloride (100 ml), water(100 ml) and a saturated aqueous
solution of
sodium hydrogen carbonate (100 ml). The organic layer was dried over magnesium
sulfate.
The solvent was removed in vacuo to give 48.24 g of crude 2-oxo-1-
azaspiro[3.3]heptane-1-
carboxylic acid tent-butylester, which was used for the next step without
purification.
'H-NMR (CDCI3): d 1.55 (s, 9 H); 1.78 (m, 1 H); 1.92 (m, 1 H); 2.18 (m, 2 H);
2.90 (m, 2 H);
3.04 {s, 1 H).
(1-(tart-Butoxycarbonylamino)cyclobutyl)acetic acid:
H C CH3 O O
H3C- _O_ _N OH
3 H
An 1 N aqueous solution of lithium hydroxide (227 ml, 227 mmol) was added to a
solution of
2-oxo-1-azaspiro[3.3]heptane-1-carboxylic acid tent-butylester (48 g, 0.227
mmol) in tetrahy-
drofuran (200 ml). The reaction mixture was stirred for 2 h. Diethyl ether
(200 ml) and water
(200 ml) were added. The mixture was stirred for 16 h. The organic layer was
isolated. The
aqueous phase was extracted with diethyl ether (200 ml). The aqueous phase was
acidified
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with a 10% aqueous solution of sodium hydrogen sulfate until pH 3. The formed
precipitation
was filtered off, washed with water, and dried in vacuo, to give 38.84 g of (1-
(tert-
butoxycarbonylamino)cyclobutyl)acetic acid.
'H-NMR (CDCI3): d 1.45 (s, 9 H); 1.85 (m, 1 H); 1.95 (m, 1 H); 2.25 (m, 4 H);
2.87 (m, 2 H);
5.15 and 6.20 (both br, together 1 H).
b
(2E~-4-(1-(tent-Butoxycarbonylamino)cyclobutyl)but-2-enoic acid:
H C CH3 O O
H3C~O~N \ OH
3 H
(2~-4-(1-(tent-Butoxycarbonyiamino)cyclobutyl)but-2-enoic acid was synthesized
starting
with (1-(tent-butoxycarbonylamino)cyclobutyl)acetic acid analogously to the
synthesis of (2E)-
5-(tert-butoxycarbonylamino)-5-methylhex-2-enoic acid starting with of 3-tert-
butoxycarbonyl-
amino-3-methylbutanoic acid.
'H-NMR (CDCI3): d 1.43 (s, 9 H); 1.84 (m, 1 H); 1.95 (m, 1 H); 2.10 (m, 2 H);
2.20 (m, 2 H);
2.70 (m, 2 H); 4.75 (br, 0.5 H); 5.90 (m, 1 H); 6.35 (br, 0.5 H); 6.95 (m, 1
H).
(2E)-5-tent-Butoxycarbonylamino-3,5-dimethylhex-2-enoic acid ethyl ester.
H C CH3 O H3C CH3 CH3 O
H3C_ _O_ _N \ O~CH
3 H 3
Diacetonamine hydrogen oxalate (30.0 g; 146 mmol) was suspended in
tetrahydrofuran (400
ml). An aqueous solution of sodium hydroxide (1 N; 146 ml) was added. Di-tert-
butyl
dicarbonate (38.3 g; 175 mmol) was dissolved in tetrahydrofuran (100 ml) and
added dropwise
to the reaction mixture. The reaction mixture was stirred for 2 h at room
temperature. Sodium
hydroxide (1 N; 146 ml) was added and the reaction mixture was stirred for 12
h at room
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temperature. Water (200 ml) and ethyl acetate (200 ml) were added. The aqueous
phase was
extracted with ethyl acetate (4 x 200 ml). The combined organic phases were
dried over
magnesium sulfate, and the solvent was removed in vacuo. The residue was
purified by flash
chromatography on silica (200 g), using ethyl acetate/heptane (1:3) as eluent,
to afford 28.4 g
of (1,1-dimethyl-3-oxobutyl)carbamic acid tent-butyl ester.
Triethyl phosphonoacetate (4.7 g; 20.9 mmol) was dissolved in tetrahydrofuran
(36 ml).
Potassium tent butoxide (2.3 g; 20.9 mmol) was added and the reaction mixture
was stirred for
40 min at room temperature. (1,1-Dimethyl-3-oxobutyl)carbamic acid tent-butyl
ester (2.5 g;
11.6 mmol) was dissolved in tetrahydrofuran (15 ml) and added dropwise to the
reaction
mixture which was heated to reflux for 12 h. Ethyl acetate (100 ml) and
hydrochloric acid (1 N;
100 ml) were added and the phases were separated. The aqueous phase was
extracted with
ethyl acetate (3 x 50 ml). The combined organic phases were washed with an
aqueous solution
of sodium hydrogen carbonate (saturated; 100 ml), dried (magnesium sulfate)
and evaporated
in vacuo. The residue was purled by flash chromatography on silica (120 g)
using ethyl
acetate/heptane (1:2) as eluent to afford 2.0 g of (2E)-5-tent-
butoxycarbonyiamino-3,5-
dimethylhex-2-enoic acid ethyl ester.
'H-NMR (CDCl3) d 1.25 (t, 3H); 1.30 (s, 6H); 1.44 (s, 9H); 2.21 (s, 3H); 2.58
(s, 2H); 4.14 (q,
2H); 4.48 (s, 1 H); 5.fi5 (s, 1 H).
(2E)-5-tert-Butoxycarbonylamino-3,5-dimethylhex-2-enoic acid:
H C CH3 O H3C CH3 CH3 O
H3CI 'O- _N ~ OH
3 H
(2E)-5-tent Butoxycarbonylamino-3,5-dimethylhex-2-enoic acid ethyl ester (1.95
g; 6.83 mmol)
was dissolved in 1,4-dioxane (25 ml) and water (15 ml). Lithium hydroxide
(0.18 g; 7.52 mmol)
was added and the reaction mixture was stirred for 12 h at room temperature.
Water (150 ml)
and tent butyl methyl ether (150 ml) was added. The aqueous phase was diluted
with a 10%
aqueous solution of sodium hydrogensulfate until pH 2,5 and extracted with
tert-butyl methyl
ether (3 x 100 ml). The combined organic phases were dried over magnesium
sulfate and
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evaporated in vacuo. The residue was recrystallized from heptane (20 ml) to
afford 0.6 g of
(2E)-5-tent-Butoxycarbonylamino-3,5-dimethylhex-2-enoic acid.
'H-NMR (CDCI3) d 1.29 (s, 6H); 1.44 (s, 9H); 2.23 (s, 3H); 2.62 (s, 2H); 4.45
(s, 1H); 5.66 (s,
1 H).
(2E)-5-(N-(tert Butoxycarbonyl)-N-methylamino)-5-methylhex-2-enoic acid.
H C CH3 O H3C CH3 O
H3C_ _O- _N \ OH
3 I
CH3
(2E)-5-(tert-Butyloxycarbonylamino)-5-methylhex-2-enoic acid (5.00 g ; 20.6
mmol) was
dissolved in tetrahydrofuran (70 ml). Methyliodide (10.3 ml; 164 mmol) was
added and the
solution was cooled to 0° C. Sodium hydride (60% in oil)( 2.07 g; 61.6
mmol) was added in
portions and the solution was stirred at roomtemperature for four days. Ethyl
acetate (70 ml)
and water (60 ml) was added dropwise and the solvent was removed in vacuo. The
crude
product was dissolved in water (40 ml) and ether (40 ml). The organic phase
was washed
with a saturated aqueous solution of sodium hydrogencarbonate (30 ml). The
aqueous
phases were mixed and 5% aqueous citric acid was added to pH 3. The aqueous
phase was
extracted with ethylacetate (4 x 50 ml). The organic phase was washed with
water (2 x 40
ml), an aqueous solution of sodium thiosulfate (5%; 40 ml}, water (40 ml),
dried over MgS04
and the solvent was removed in vacuo. The residue was dissolved in
ethyiacetate (45 ml)
and washed with an aqueous solution of sodium hydrogensuifate (10%; 3 x 30
ml), dried
over MgSO, and and concentrated in vacuo to give 4.00 g of (2E)-5-(N-(tert
Butoxy-
carbonyl}-N-methylamino)-5-methylhex-2-enoic acid.
'H-NMR (CDC13) b 1.38 (s, 6H), 1.45 (s, 9H ); 2.80 (d, 2H); 2.85 (s, 3H); 5.88
(d, 1 H); 7.01
(q, 1 H ).
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Example 1
(2E)-5-Amino-5-methylhex-2-enoic acid N-((1R)-1-{N-[(1R)-1-benzyl-2-(4-
((dimethylamino)methyl)piperidin-1-yl)-2-oxoethylJ-N-methylcarbamoyl}-2-(2-
naphthyl)ethyl)-
5 N-methylamide
H3C CH3 O CH3 O
H2N ~ N~ N N CH3
I II I
CH3 O NCH
3
4-(Dimethylcarbamoyl)piperidine-1-carboxylic acid tert-butyl ester
CH3 O
HC- 1_ - _N CH
3 C~ N 3
CH3
O
1-(tert-Butoxycarbonyl)piperidine-4-carboxylic acid (8.0 g, 35 mmol) was
dissolved in di-
chloromethane (70 ml) and N,N-dimethylformamide (35 ml). 1-Hydroxy-7-
azabenzotriazole
(4.75 g, 35 mmol) was added. The solution was cooled to 0 °C. N-(3-
dimethylaminopropyl)-
N'-ethylcarbodiimide hydrochloride (6.69 g, 35 mmol) was added. The reaction
mixture was
stirred for 20 min at 0 °C. A 5.6 M solution of dimethylamine in
ethanol (37 ml, 209 mmo!)
was added. The reaction mixture was stirred for 3 days, while it was warming
up to room
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5s
b
temperature. It was diluted with ethyl acetate (400 ml) and washed with a 10%
aqueous so-
lutiorr~of sodium hydrogen sulfate (400 ml). The aqueous phase was extracted
with ethyl
acetate (2 x 200 ml). The combined organic layers were washed with a saturated
aqueous
solution of sodium hydrogen carbonate (300 ml) and dried over magnesium
sulfate. The sol-
ven was removed in vacuo. The crude product was purified by flash
chromatography on sil-
ica (300 g), using dichloromethane/methanol 20:1 as eluent, to give 4.56 g of
4-(dimethyl-
carbamoyl)piperidine-1-carboxylic acid tert-butyl ester.
'H-NMR (CDCl3): s 1.47 (s, 9 H); 1.70 (m, 4 H); 2.60 - 2.90 (m, 3 H); 2.96 (s,
3 H); 3.08 (s, 3
H); 4.17 (m, 2 H).
4-((Dimethylamino)methyf)piperidine-1-carboxylic acid tert-butyl ester
CH3 O
H CI , _O_ _N CH
3 CH3 I 3
N~CH3
At 0 °C a solution of 4-((dimethylamino)methyl)piperidine-1-carboxylic
acid tent-butyl ester
(4.56 g, 18 mmol) in tetrahyrofuran (80 ml) was added dropwise to a suspension
of sodium
borohydride (1.61 g, 43 mmol) in tetrahydrofuran (80 ml). The reaction mixture
was stirred
for 20 min at 0 °C. A solution of iodine 4.51 g, 18 mmoi) in
tetrahydrofuran (80 ml) was
added dropwise at 0 °C. The reaction mixture was heated to reflux for
16 h. It was cooled to
4 °C. Methanol (200 ml) was added dropwise. The solvent was removed in
vacuo. The resi-
due was dissolved in a 20% aqueous solution of sodium hydroxide (200 ml) and
tert-butyl
methyl ether (150 ml). The phases were separated. The aqueous phase was
extracted with
tert-butyl methyl ether (3 x 100 ml). The combined organic layers were dried
over magne-
sium sulfate. The solvent was removed in vacuo. The crude product was purified
by flash
chromatography on silica (100 g), using dichloromethane/methanoll25% aqueous
ammonia
(100:10:1) as eluent, to give 4.07 g of 4-((dimethylamino)methyl)piperidine-1-
carboxylic acid
tert-butyl ester.
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'H-NMR (CDC13): 8 1.22 (m, 2 H); 1.44 (s, 9 H); 1.85 (d, 2 H); 2.09 (m, 1 H);
2.61 (s, 6 H);
2.65 (m, 2 H); 2.78 {t, 2 H); 4.05 (d, 2 H).
N,N-Dimethyl-N-((piperidin-4-yl)methyl)amine
CH3
N~CH3
H
A 3 M solution of hydrogen chloride in ethyl acetate (120 ml, 360 mmol) was
added to a so-
lution of 4-((dimethylamino)methyl)piperidine-1-carboxylic acid tert-butyl
ester {2.0 g, 14
mmol) in ethyl acetate (50 ml). The reaction mixture was stirred for 30 min at
room tem-
peratufe. The solvent was removed in vacuo to give 2.3 g of the crude
dihydrochloride salt of
N,N-dimethyl-N-((piperidin-4-yl)methyl)amine, which was used without
purification for the
next step.
'H-NMR (CDCI3, selected values): b 1.48 (m, 2 H); 1.92 (s, 6 H); 3.22 (d, 2
H).
N-[(1 R)-1-Benzyl-2-(4-((dimethylamino)methyl)piperidin-1-yl)-2-oxoethyl]-N-
methylcarbamic
acid tert-butyl ester
CH3 O N~CH3
~ ~ N~ CH3
H CI , _O_ _N
~-13C
CH3 O
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At 0 °C, N-(3-dimethylaminopropyl}-N'-ethylcarbodiimide hydrochloride
(1.158 g, 6.04 mmol)
was added to a solution of (2R}-2-(N-(tert-butoxycarbonyl)-N-methylamino)-3-
phenylpropionic acid (1-69 g, 6.04 mmol) and 1-hydroxy-7-azabenzotriazole
(0.822 g, 6.04
mmol) in dichloromethane (25 ml) and N,N-dimethylformamide (12 ml). The
reaction mixture
was stirred for 20 min at 0 °C. A solution of the crude dihydrochloride
salt of N,N-dimethyl-N-
((piperidin-4-yl)methyl)amine (1.3 g, 6.04 mmol) in N,N-dimethylformamide (10
ml) and di-
chloromethane (5 ml) and ethyldiisopropylamine (6.2 ml, 36.25 mmol) were added
succes-
sively. The reaction mixutre was stirred for 16 h, while it was warming up to
room tempera-
ture. It was diluted with ethyl acetate (100 ml) and washed with a saturated
aqueous solution
of sodium hydrogen carbonate (100 ml). The organic layer was dried over
magnesium sul-
fate. The solvent was removed in vacuo. The crude product was purified by
falsh chromatog-
raphy on silica (100 g), using dichloromethane/methanoU25% aqueous ammonia
(200:10:1 )
as eluent, to give 1.22 g of N-[(1 R)-1-benzyl-2-(4-
((dimethylamino)methyl)piperidin-1-yl)-2-
oxoethyl]-N-methylcarbamic acid tert-butyl ester.
'H-NMR (CDCI3, selected values): & 1.28, 1.11, 1.37, and 1.38 (all s, together
9 H); 4.00 (m,
1 H); 4.57 (m, 1 H); 4.97 and 5.28 (both t, together 1 H); 7.10 - 7.40 (m, 5
H).
MS: 404 [M+1 )~
(2R}-1-(4-((Dimethylamino)methyl)piperidin-1-yl)-2-(methylamino)-3-
phenylpropan-1-one
N~CH3
N J CH3
HN
CH3 I IO
At 0 °C, trifluoroacetic acid (20 ml) was added to a solution of N-
[(1R)-1-benzyl-2-(4-((di-
methylamino)methyl)piperidin-1-yl)-2-oxoethyl]-N-methylcarbamic acid tert-
butyl ester (1.22
g, 3.02 mmol) in dichloromethane (20 ml). The reaction mixture was stirred for
1 h at 0 °C.
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The solvents were removed in vacuo. The residue was dissolved in
dichloromethane (70 ml)
and the solvent was removed in vacuo. The latter procedure was repeated two
times. The
crude product was purified by flash chromatography on silica (100 g), using
dichloro-
methane/methanol/25% aqueous ammonia (100:10:1 ), to give 659 mg of (2R)-1-(4-
((dimethylamino)methyl)piperidin-1-yl)-2-(methylamino)-3-phenylpropan-1-one.
'H-NMR (CDCI3, selected values): 8 0.91 and 1.47 (m and d, together 1 H); 1.27
(m, 1 H);
4.62 (t, 1 H).
N-((1R}-1-{N-[(1R)-1-Benzyl-2-(4-((dimethylamino)methyl)piperidin-1-yl)-2-
oxoethylJ-N-
methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-methylcarbamic acid tert-butyl ester
1
CH3 O CH3 O
H C' / _O- _N N N CH
3hi3C I ~ I
CH3 O NCH
3
At 0 °C, N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride
(416 mg, 2.17 mmol)
was added to a solution of (2R)-2-(N-(tert-butoxycarbonyl)-N-methylamino)-3-(2-
naphthyl)-
propioni acid (715 mg, 2.17 mmol) and 1-hydroxy-7-azabenzotriazole (296 mg,
2.17 mmol)
in dichloromethane (20 ml) and N,N-dimethylformamide (10 ml). The reaction
mixture was
stirred for 20 min at 0 °c. A solution of (2R)-1-(4-
((dimethylamino)methyl)piperidin-1-yl)-2-
(methylamino)-3-phenylpropan-1-one (659 mg, 2.17 mmol) in dichloromethane (10
ml) and
N,N-dimethylformamide (5 ml) and ethyldiisopropylamine (0.56 ml, 3.26 mmol)
were added
successively. The reaction mixture was stirred for 16 h, while it was warming
up to room
temperature. It was diluted with ethyl acetate (100 ml) and washed with a
saturated aqueous
solution of sodium hydrogen carbonate (100 ml). The aqueous solution was
extracted with
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ethyl acetate (3 x 50 ml). The combined organic layers were dried over
magnesium sulfate.
The solvent was removed in vacuo. The crude product was purified by flash
chromatography
on silica (80 g), using ethyl acetate/heptane/triethylamine (1:1:0.08) as
eluent, to give 1.05 g
of N-((1R)-1-{N-[(1R)-1-benzyl-2-(4-((dimethylamino)methyl)piperidin-1-yl)-2-
oxoethylj-N-
5 methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-methylcarbamic acid tent-butyl
ester.
'H-NMR (CDCl3, selected values): b 1.24 and 1.42 (both s, together 9 H); 5.04,
5.28, 5.44,
5.54, 5.73 (m, dd, dd, dd, and m, together 3 H);
(2R)-N-[(1 R)-1-Benzyl-2-(4-((dimethylamino)methyl)piperidin-1-yl)-2-oxoethyl]-
N-methyl-2-
(methylamino)-3-{2-naphthyl)propionamide
CH3 O
HN~ N N CH3
I II I
CH3 O NCH
3
At 0 °C, trifiuoroacetic acid (18 ml) was added to a solution of N-
((1R)-1-{N-[(1R)-1-benzyl-2-
(4-((dimethylamino)methyl)piperidin-1-yl)-2-oxoethylj-N-methylcarbamoyl}-2-(2-
naphthyl)-
ethyl}-N-methylcarbamic acid tert-butyl ester (1.05 g, 1.71 mmol) in
dichloromethane (18 ml).
The reaction mixture was stirred for 50 min at 0 °C. The solvents were
removed in vacuo.
The residue was dissolved in dichloromethane (50 ml) and the solvent was
removed in
vacuo. The latter procedure was repeated two times. The crude product was
purified by flash
chromatography on silica (80 g), using dichloromethane/methanol/25% aqueous
ammonia
as eluent, to give 846 mg of (2R)-N-[(1R)-1-benzyl-2-(4-
((dimethylamino)methyl)piperidin-1-
yl)-2-oxoethylJ-N-methyl-2-(methylamino}-3-{2-naphthyl)propionamide.
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'H-NMR (CDC13, selected values): 8 0.60 (m, 1 H); 4.38 (t, 1 H); 5.72 and 5.79
(both t, to-
gether 1 H).
{(3E)-4-[N-{(1 R)-1-{N-[(1 R)-1-Benzyl-2-(4-((dimethylamino)methyl)piperidin-1-
yl)-2-oxoethyl]-
N-methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-methylcarbamoyl]-1,1-dimethylbut-3-
enyl}carbamic acid tert-butyl ester
CH3 O H3C CH3 O CH3 O
H C_ 1 _~~_ _N ~ N N N CH
C 1"13 H I
CH3 O NCH
3
At 0 °C, N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride
(112 mg, 0.58 mmol)
was added to a solution of (2E)-5-(tert-butoxycarbonylamino)-5-methylhex-2-
enoic acid (142
mg, 0.58 mmol) and 1-hydroxy-7-azabenzotriazole (79 mg, 0.58 mmol) in
dichloromethane
(10 ml) and N,N-dimethylformamide (5 ml). The reaction mixture was stirred for
20 min at 0
°C. A solution of (2R)-N-[(1 R)-1-benzyl-2-(4-
((dimethylamino)methyl)piperidin-1-yl)-2-
oxoethylj-N-methyl-2-(methylamino)-3-(2-naphthyl)propionamide (300 mg, 0.58
mmol) in di-
chloromethane (5 ml) and N,N-dimethylformamide (5 ml) and
ethyldiisopropylamine 0.10 ml,
0.58 mmol) were added successively. The reaction mixture was stirred for 3
days, while it
was warming up to room temperature. It was diluted with ethyl acetate (70 ml)
and washed
with a saturated aqueous solution of sodium hydrogen carbonate (70 ml). The
aqueous
phase was extracted with ethyl acetate (3 x 50 ml). The combined organic
layers were dried
over magnesium sulfate. The solvent was removed in vacuo. The crude product
was purified
by flash chromatography on silica (70 g), using dichloromethanelmethanol/25%
aqueous
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ammonia (200:10:1) as eluent, to give 313 g of {(3E)-4-[N-((1 R)-1-{N-[(1 R)-1-
benzyl-2-(4-
((dimethylamino)methyl)piperidin-1-yl)-2-oxoethyl]-N-methylcarbamoyl}-2-(2-
naphthyl)ethyl)-
N-methylcarbamoyl]-1,1-dimethylbut-3-enyl}carbamic acid tert-butyl ester.
'H-NMR (CDCI3, selected values): b 1.28 and 1.30 (both s, together 6 H); 1.42
(s, 9H); 2.23,
2.27, 2.38, 2.43, 2.51, 2.52, 2.81, and 2.82 (all s, together 12 H); 5.56,
5.76, and 5.90 (m, m,
and dd, together 2 H); 6.17 and 6.19 (both dd, together 1 H); 6.94 (m, 1 H).
At 0 °C, trifluoroacetic acid (6 ml) was added to a solution of {(3E)-4-
[N-((1 R)-1-{N-[(1 R)-1-
benzyl-2-(4-((dimethylamino)methyl)piperidin-1-yl)-2-oxoethyl]-N-
methylcarbamoyl}-2-(2-
naphthyl)ethyl)-N-methyicarbamoyl]-1,1-dimethylbut-3-enyl}carbamic acid tert-
butyl ester
(212 mg, 0.29 mmol) in dichloromethane {6 ml). The reaction mixture was
stirred for 20 min
at 0 °C. It was diluted with dichloromethane (30 ml). A saturated
aqueous solution of sodium
hydrogen carbonate (30 ml) was added dropwise. Solid sodium hdyrogen carbonate
was
added, until pH 7 was obtained. The phases were separated. The aqueous phase
was ex-
tracted with dichloromethane (3 x 50 ml). The combined organic layers were
dried over
magnesium sulfate. The solvent was removed in vacuo. The crude product was
purified by
flash chromatography on silica (20 g), using dichloromethane/methanol/25%
aqueous am-
monia (100:10:1 ) as eluent, to give 5 mg of the title compound.
'H-NMR (CDCI3, selected values): 8 1.20 {s, 6 H); 2.28, 2.32, 2.41, 2.49,
2.56, 2.57, 2.82,
and 2.83 (all s, together 12 H); 5.58, 5.78, and, 5.92 (m, m, and dd, together
2 H); 6.16 and
6.19 (both d, together 1 H); 7.00 (m, 1 H).
HPLC: 39.23 min (A1 ).
41.55 min (B1).
MS: 640.4 [M+1 ]'.
For biological testing, the title compound was transferred into its acetate
salt by lyophilization
with 0.5 M acetic acid (40 ml).
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Example 2
(2E)-5-Amino-5-methylhex-2-enoic acid N-((1 R)-1-{N-[(1 R)-1-benzyl-2-((3S)-3-
(dimethylaminomethyl)piperidin-1-yl)-2-oxoethyl]-N-methylcarbamoyl}-2-(2-
naphthyl)ethyl)-N-
methylamide
/ /
H3C CH3 O CH3 O
H2N ~ N~ N N
I II
CH3 O
\ N.CHs
r I
CH3
't 0
(3R)-Piperidine-1,3-dicarboxylic acid 1-tert-butyl ester 3-ethyl ester
O
O~CH3
N~
O- ' O
H C- I _CH
3 CH3 3
(R)-Ethyl nipetcotate tartrate (10.0 g, 32.5 mmol) were suspended in
tetrahydrofuran {90 ml).
An 1 N solution of sodium hydroxide in water (98 ml, 98 mmol) was added. A
solution of di-
tert-butyl Bicarbonate (7.10 g, 32.5 mmol) in tetrahydrofuran (90 ml) was
added. The reac-
tion mixture was stirred for 16 h at room temperature. Ethyl acetate (400 ml)
was added. The
reaction mixture was washed with a 10% aqueous solution of sodium hydrogen
sulfate (400
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ml). The aqueous solution was extracted with ethyl acetate ( 2 x 200 ml). The
combined or-
ganic layers were washed with a saturated aqueous solution of sodium hydrogen
carbonate
(200 ml) and dried over magnesium sulfate. The solvent was removed in vacuo.
The crude
product was purified by flash chromatography on silica (90 g), using ethyl
acetate/heptane
1:4 as eluent, to give 4.13 g of (3R)-piperidine-1,3-dicarboxylic acid 1-tert-
butyl ester 3-ethyl
w
ester.
'H-NMR (CDCl3): s 1.27 (t, 3 H); 1.48 (s, 9 H); 1.54 (m, 1 H); 1.62 {m, 1 H);
1.73 (m, 2H);
2.05 (m, 1 H); 2.45 (m, 1 H); 2.81 (m, 1 H); 2.98 (br, 1 H); 3.93 (m, 1 H);
4.14 (q, 1 H).
(3R)-3-Formylpiperidine-1-carboxylic acid tert-butyl ester
O
~H
N
O_ ' O
H C_ I _CH
3 CH3 3
A 1.2 M solution of diisobutylaiuminum hydride in toluene (30.8 ml, 36.9 mmol)
was added at
-78 °C to a solution of (3R)-piperidine-1,3-dicarboxylic acid 1-tent-
butyl ester 3-ethyl ester
(4.13 g, 16.1 mmol) in diethyl ether (30 ml). The reaction mixture was stirred
for 2.5 h at - 78
°C. Water (9.6 ml) was added dropwise. The reaction mixture was warmed
to room tem-
perature. The precipitation was removed by filtration through a plug of
celite. The celite was
washed with tert-butyl methyl ether (3 x 100 ml). The liquids were combined
and dried over
magnesium sulfate. The solvent was removed in vacuo, to give 1.94 g of crude
(3R)-3-
formylpiperidine-1-carboxylic acid tert-butyl ester, which was used for the
next step without
further purification.
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'H-NMR (CDC13): 8 1.45 (s, 9H); 1.67 (m, 2 H); 1.95 (m, 1 H); 2.43 (m, 1 H);
3.10 (m, 1 H);
3.32 (dd, 1 H); 3.52 (d, 1 H); 3.66 (m, 1 H); 3.95 (m, 1 H); 9.69 (s, 1 H).
5 (3S)-3-(Dimethylaminomethyl)piperidine-1-carboxylic acid tert-butyl ester
O- ' O
H C_ I -CH
3 CH3 3
A solution of crude (3R)-3-formylpiperidine-1-carboxylic acid tert-butyl ester
(1.94 g, 9.1
10 mmol) in dichloromethane (80 ml) was prepared. A 5.6 M solution of
dimethylamine in etha-
nol (3.2 ml, 18.2 mmol) and molsieves were added successively. Sodium
triacetoxyborohy-
dride (5.78 g, 27.3 mmol) was added to this mixture. Acetic acid (1.04 ml,
18.2 mmol) was
added. The reaction mixture was stirred for 16 h at room temperature. An 1 N
aqueous solu-
tion of sodium hydroxide (70 ml) and tert-butyl methyl ether (70 ml) were
added. The phases
15 were separated. The aqueous solution was extracted with tert-butyl methyl
ether (3 x 70 ml).
The combined organic layers were dried over magnesium sulfate. The solvent was
removed
in vacuo. The crude product was purified by flash chromatography on silica (40
g), using di-
chloromethanelmethanoll25% aqueous ammonia (100:10:1) as eluent, to give 866
mg of
(3S}-3-(dimethylaminomethyl)piperidine-1-carboxylic acid tert-butyl ester.
N~CH3
J ~H3
N
'H-NMR (CDC13): b 1.10 (m, 1 H); 1.45 (s, 9 H), 1.45 {m, 1 H); 1.64 (m, 2 H);
1.85 (m, 1 H);
2.10 (m, 2 H); 2.20 (s, 6 H); 2.50 (br, 1 H); 2.79 (m, 1 H); 3.95 (m, 2 H).
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N, N-Dimethyl-N-(((3R)-piperidin-3-yl)methyl)amine
N~CH3
N J cH3
H
(3S)-3-(Dimethylaminomethyl)piperidine-1-carboxylic acid tert-butyl ester
(1.25 g, 5.15
mmol) was dissolved in ethyl acetate (30 ml). A 2.7 M solution of hdyrogen
chloride in ethyl
acetate (75 ml, 203 mmol) was added. The reaction mixture was stirred for 45
min at room
temperature. The solvent was removed in vauco to give 976 mg of the crude
dihydrochloride
salt of N,N-dimethyl-N-(((3R)-piperidin-3-yl)methyl)amine, which was used for
the next step
without further purification.
20
'H-NMR (CD30D): S 1.42 (m, 1 H); 1.86 (m, 1 H); 2.00 (m, 2 H); 2.38 (m, 1 H);
2.85 (t, 1 H);
2.95 (s, 6 H); 2.98 (m, 1 H); 3.16 (m, 2 H); 3.42 (m, 1 H); 3.53 (m, 1 H).
N-[(1 R)-1-Benzyl-2-((3S)-3-(dimethylaminomethyl)piperidin-1-yl)-2-oxoethyl]-N-
methylcarbamic acid tert-butyl ester
CH3 O
~N
H3C' I O I I N
CH3 O
N~CH3
I
CH3
At 0 °C N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride
(870 mg, 4.54 mmol)
was added to a solution (2R)-2-(N-(tert-butoxycarbonyl)-N-methyiamino)-3-
phenylpropionic
acid (1.27 g, 4.54 mmol} and 1-hydroxy-7-azabenzotriazole (617 mg, 4.54 mmol)
in dichloro-
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methane (20 ml) and N,N-dimethylformamide (10 ml). The reaction mixture was
stirred for 20
min at 0 °C. A solution of the crude dihydrochloride salt of N,N-
tiimethyl-N-(((3R)-piperidin-3-
yl)methyl)amine (976 mg, 4.54 mmol) in dichloromethane (20 ml) and N,N-
dimethyl-
formamide (10 ml) and ethyldiisopropylamine (3.9 ml, 22.7 mmol) were added
successively.
The reaction mixture was stirred for 3 days, while it was warming up to room
temperature.
Ethyl acetate (300 ml) was added. The solution was washed with a saturated
aqueous solu-
tion of sodium hydrogen carbonate (300 ml). The aqueous phase was extracted
with ethyl
acetate (2 x 200 ml). The combined organic layers were dried over magnesium
sulfate. The
solvent was removed in vacuo. The crude product was purified by flash
chromatography on
.. 10 silica (90 g), using dichloromethanelmethanoll25% aqueous ammonia
(100:10:1) as eluent,
to give 1.69 g of N-[(1R)-1-benzyl-2-((3S)-3-(dimethylaminomethyl)piperidin-1-
yl)-2-
oxoethylJ-N-methylcarbamic acid tert-butyl ester.
'H-NMR (CDCI3, selected values): 8 1.20, 1.24, 1.31, and 1.32 (all s, together
9 H); 2.12,
2.13, and 2.18 (all s, together 6 H); 2.81 (m, 3 H); 4.97 and 5.30 (both m,
together 1 H); 7.05
- 7.35 (m, 5 H).
(2R)-1-((3S)-3-((Dimethylamino)methyl)piperidin-1-yl)-2-methylamino-3-
phenylpropan-1-one
CH3 O
HN
~N
~ N~CH3
I
CH3
At 0 °C, trifluoroacetic acid (25 ml) was added to a solution of N-
[(1R)-1-benzyl-2-((3S)-3-
(dimethylaminomethyl)piperidin-1-yl)-2-oxoethyl]-N-methylcarbamic acid tert-
butyl ester (1.69
g, 4.2 mmol) in dichloromethane (25 ml). The reaction mixture was stirred for
30 min at 0 °C.
The solvent was removed in vacuo. The residue was dissolved in dichioromethane
(100 ml)
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and the solvent was removed in vacuo. The latter procedure was repeated two
times. The
crude product was purified by flash chromatography on silica (90 g), using
dichloro-
methane/methanol/25% aqueous ammonia (100:10:1) as eluent, to give 1.15 g of
(2R)-1-
((3S)-3-((dimethylamino)methyl)piperidin-1-yl}-2-methylamino-3-phenylpropan-1-
one.
'H-NMR (CDCl3, selected values): 8 0.38, 1.11, 1.37, and 1.65 (all m, together
4 H); 2.11,
2.19, 2.25, and 2.31 (all s, together 9 H); 4.37 and 4.53 (both m, together 1
H); 7.10 - 7.35
(m, 5 H).
N-(( 1 R)-1-{N-[( 1 R)-1-Benzyl-2-((3S)-3-((dimethylamino)methyl)piperidin-1-
yl)-2-oxoethylj-N-
methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-methylcarbamic acid tert-butyl ester
H C CH3 O CH3 O
H3C' _O_ _N~ N N
I II
CH3 O
N~CH3
I
CH3
At 0 °C N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride
(379 mg, 1.98 mmol)
was added to a solution of (2R)-2-(N-(tert-butoxycarbonyl}-N-methylamino)-3-(2-
naphthyl)-
propionic acid (651 mg, 1.98 mmol) and 1-hydroxy-7-azabenzotriazoie (269 mg,
1.98 mmol)
in dichloromethane (10 ml) and N,N-dimethylformamide (5 ml). The reaction
mixture was
stirred for 20 min at 0 °C. A solution of (2R)-1-((3S)-3-
((dimethylamino)methyl)piperidin-1-yl)-
2-methylamino-3-phenylpropan-1-one (600 mg, 1.98 mmol) in dichloromethane (10
ml) and
ethyldiisopropylamine (0.51 ml, 2.97 mmol) were added successively. The
reaction mixture
was stirred for 3 days, while it was warming up to room temperature. Ethyl
acetate (100 ml)
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was added. The solution was washed with a saturated aqueous solution of sodium
hydrogen
carbonate (100 ml). The aqueous phase was extracted with ethyl acetate (3 x 50
ml). The
combined organic layers were dried over magnesium sulfate. The solvent was
removed in
vacuo. The crude product was purified by flash chromatography on silica (90
g), using di-
chloromethanelmethanoU25% aqueous ammonia (100:10:1) as eluent, to give 1.18 g
of N-
((1 R)-1-{N-[(1 R)-1-benzyl-2-((3S)-3-((dimethylamino)methyl)piperidin-1-yl)-2-
oxoethylj-N-
methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-methylcarbamic acid tert-butyl ester.
'H-NMR (CDCl3, selected values): 8 0.45 and 0.71 (both m, together 1 H); 1.03,
1.05, 1.15,
1.20, 1.28, 1.36, and 1.42 (all s, together 9 H); 2.12, 2.15, 2.21, 2.26,
2.29, 2.85 (all s, to-
gether 6 H); 5.05, 5.44, 5.58, 5.71, 5.85, and 6.00 (all s, together 2 H);
7.10 - 7.80 (m, 12 H).
(2R)-N-[(1 R)-1-Benzyl-2-((3S)-3-((dimethyiamino)methyl)piperidin-1-yl)-2-
oxoethylj-N-
methyl-2-(methylamino)-3-(2-naphthyl)propionamide
/ /
CH3 O
w. N
HN~ N
CH3 ' IO
~ N.CHs
'- I
CH3
2o At 0 °C, trifluoroacetic acid (20 ml) was added to a solution of N-
((1R)-1-{N-[(1R)-1-benzyl-2-
((3S)-3-((dimethylamino)methyl)piperidin-1-yl)-2-oxoethylj-N-methylcarbamoyl}-
2-(2-
naphthyl)ethyl)-N-methylcarbamic acid tert-butyl ester (1.188, 1.92 mmol) in
dichloro-
methane (20 ml). The reaction mixture was stirred for 50 min at 0 °C.
The solvent was re-
moved in vacuo. The residue was dissolved in dichloromethane (80 ml) and the
solvent was
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removed in vacuo. The latter procedure was repeated two times. The crude
product was pu-
rified by flash chromatography on silica (40 g), using
dichloromethane/methanol/25% aque-
ous ammonia (100:10:1) as eluent, to give 788 mg of (2R)-N-[(1R}-1-benzyl-2-
((3S)-3-
((dimethylamino)methyl)piperidin-1-yl)-2-oxoethyl}-N-methyl-2-(methylamino)-3-
(2-naphthyl)-
propionamide.
'H-NMR (CDCI3, selected values): 8 2.01 and 2.25 (both s, together 9 H); 3.72
(m, 2 H); 3.95
and 4.27 (both m, together 1 H); 5.77, 5.86, and 6.03 (t, m, and dd, together
1 H); 7.10 and
7.85 (m, 12 H).
{(3E)-4-[N-(( 1 R)-1-{N-[( 1 R)-1-Benzyl-2-((3S)-3-
((dimethylamino)methyl)piperidin-1-yl)-2-
oxoethyl]-N-methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-methylcarbamoyl]-1,1-
dimethylbut-3-
enyl}carbamic acid tert-butyl ester
/ /
H C CH3 OH3C CH3 O CH3 O
H3C"O"N ~ N~ N N
H I II
CH3 O
N~CH3
I
CH3
At 0 °C, N-(3-dimethylaminopropyl}-N'-ethylcarbodiimide hydrochloride
(105 mg, 0.55 mol)
was added to a solution of (2E)-5-(tert-butoxycarbonylamino)-5-methylhex-2-
enoic (136 mg,
0.55 mmol) and 1-hydroxy-7-azabenzotriazole (74 mg, 0.55 mmol) in
dichloromethane (5
ml). The reaction mixture was stirred for 20 min at 0 °C. A solution of
(2R)-N-[(1 R)-1-benzyl-
2-((3S)-3-((dimethylamino)methyl)piperidin-1-yl)-2-oxoethyl]-N-methyl-2-
(methylamino)-3-(2-
naphthyl)propionamide (281 mg, 0.55 mmol) in dichloromethane (5 ml) and N,N-
dimethylformamide (5 ml) and ethyldiisopropylamine (0.094 ml, 0.55 mmol) were
added suc-
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cessively. The reaction mixture was stirred for 16 h, while it was warming up
to room tem-
perature. It was diluted with ethyl acetate (70 ml) and washed with a
saturated aqueous so-
lution of sodium hydrogen carbonate (70 ml). The aqueous phase was extracted
with ethyl
acetate (3 x 50 ml). The combined organic layers were dried over magnesium
sulfate. The
solvent was removed in vacuo. The crude product was purified by flash
chromatography on
silica (40 g), using dichloromethanelmethanol/25% aqueous ammonia (100:10:1)
as eluent,
to give 398 mg of {(3E)-4-[N-((1 R)-1-{N-[{1 R)-1-benzyl-2-((3S)-3-
((dimethylamino)methyl)-
piperidin-1-yl)-2-oxoethyl]-N-methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-
methylcarbamoyl]-1,1-
dimethylbut-3-enyl}carbamic acid tert-butyl ester.
'H-NMR (CDC13, selected values): b 1.44 (s, 9 H); 5.58, 5.75, and 5.86 (all m,
2 H); 6.09 and
6.17 (both d, together 1 H); 6.84 (m; 1 H); 7.10 - 7.80 (m, 12 H).
At 0 °C, trifluoroacetic acid (7 ml) was added to a solution of {(3E)-4-
[N-((1R)-1-{N-((1R)-1- .
benzyl-2-((3S)-3-((dimethylamino)methyl)piperidin-1-yl)-2-oxoethyl]-N-
methylcarbamoyl}-2
(2-naphthyl)ethyl)-N-methylcarbamoyl]-1,1-dimethylbut-3-enyl}carbamic acid
tert-butyl ester
(398 mg, 0.54 mmol) in dichloromethane (7 ml). The reaction mixture was
stirred for 40 min
at 0 °C. The solvent was removed in vacuo. The residue was dissolved in
dichloromethane
(20 ml) and the solvent was removed in vacuo. The latter procedure was
repeated two times.
The crude product was purified by flash chromatography on silica (40 g), using
dichloro-
methane/methanoU25% aqueous ammonia (100:10:1) as eluent, to give 150 mg of
the title
compound.
'H-NMR (CDCI,, selected values): 8 1.08, 1.12, 1.14, and 1.15 (all s, together
6 H), 5.46,
5.59, 5.75, and 5.94 (all m, together 2 H); 6.15 (m, 1 H); 6.93 (m, 1 H).
HPLC 27.55 min (A1 ).
30.23min (B1).
LC-MS: 640.4 [M+1]' at 8.54 min.
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For biological testing, the title compound was transferred into its acetate
salt, by lyophiliza-
tion from 0.5 M acetic acid (40 ml).
Example 3
(2E)-4-(1-Aminocyclobutyl)but-2-enoic acid N-((1 R)-1-{N-[(1 R)-1-benzyl-2-
((3S)-3-(dimethyl-
aminomethyl)piperidin-1-yl)-2-oxoethyl]-N-methylcarbamoyl}-2-(2-
naphthyl)ethyl)-N-
methylamide
/ /
O CH3 O
H2N ~ N~ N N
I II
CH3 O
N~CH3
I
CH3
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(1-{(2E)-3-[N-((1R)-1-{N-[(1 R)-1-Benzyl-2-((3S}-3-
((dimethylamino)methyl)piperidin-1-yl)-2-
oxoethyl]-N-methylcarbamoyl}-2-(2-naphthyl)ethyl}-N-
methylcarbamoyl]allyl}cyclo-
butyl)carbamic acid tert-butyl ester
.
H C CH3 O O CH3 ~O
H3C' _O- _N \ N~ N N
H I II
CH3 O
. ~ ~ N~CH3
I
CH3
At 0 °C, N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride
(85 mg, 0.44 mmol)
was added to a solution of (2E}-5-(tert-butoxycarbonylamino)-5-methylhex-2-
enoic acid (113
mg, 0.44 mmol) and 1-hydroxy-7-azabenzotriazole (60 mg, 0.44 mmol) in
dichloromethane
(5 ml) and N,N-dimethylformamide (5 ml). The reaction mixture was stirred for
20 min at 0
°C. A solution of (2R)-N-[(1R)-1-benzyl-2-((3S)-3-
((dimethylamino)methyl)piperidin-1-yi)-2-
oxoethyl]-N-methyl-2-(methylamino)-3-(2-naphthyl)propionamide (228 mg, 0.44
mmol) in di-
chloromethane (10 ml) and ethyldiisopropylamine (0.07 ml, 0.44 mmol) were
added succes-
sively. The reaction mixture was stirred for 1fi h, while it was warming up to
room tempera-
ture. It was diluted with ethyl acetate (70 ml) and washed with a saturated
aqueous solution
of sodium hydrogen carbonate (70 ml). The aqueous phase was extracted with
ethyl acetate
(3 x 50 ml). The combined organic layers were dried over magnesium sulfate.
The solvent
was removed in vacuo. The crude product was purified by flash chromatography
on silica (40
g), using dichlonnethanelmethanoll25% aqueous ammonia (100:10:1 ) as eluent,
to give 314
mg of (1-{(2E)-3-[N-((1 R)-1-{N-[(1 R)-1-benzyl-2-((3S)-3-
((dimethylamino)methyl)piperidin-1-
2o yl)-2-oxoethyl]-N-methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-
methylcarbamoyl]allyf}cyclobutyl)-
carbamic acid tert-butyl ester.
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'H-NMR (CDC13, selected values): b 1.40 (m, 9 H); 4.22 and 4.67 (both m,
together 2 H);
5.60, 5.75, 5.85, and 5.90 (dd, dd, m, and m, together 2 H); 6.10 and 6.19
(both d, together
1 H); 6.73 and 6.87 (both m, together 1 H); 7.22, 7,42, and 7.76 (all m,
together 12 H).
At 0 °C, trifluoroacetic acid (7 ml) was added to a solution of (1-
{(2E}-3-[N-((1 R}-1-{N-[(1 R)-
1-benzyl-2-((3S)-3-((dimethylamino)methyl)piperidin-1-yl)-2-oxoethyl]-N-
methylcarbamoyi}-2-
(2-naphthyl)ethyl}-N-methylcarbamoyl]allyl}cyciobutyl)carbamic acid tert-butyl
ester (314 mg,
0.42 mmol) in dichlormethane (7 ml). The reaction mixture was stirred for 20
min at 0 °C.
The solvent was removed in vacuo without heating. The residue was dissolved in
dichloro-
methane (20 ml) and the solvent was removed in vacuo. The latter procedure was
repeated
two times. The crude product was purified by flash chromatography on silica
(40 g), using
dichloromethane/methanollammonia (100:10:1 ) as eluent, to give 180 mg of the
title com-
pound.
'H-NMR (CDC13, selected values): 8 0.40 and 0.74 (both m, together 2 H); 3.73
and 4.22
(both m, together 2 H); 5.57, 5.77, and 5.91 (all m, together 2 H); 6.15 and
6.24 (both d, to-
gether 1 H); 6.85 and 6.96 (both m, together 1 H); 7,22, 7,92 , and 7.74 (all
m, together 12
H).
HPLC: 28.03 min (A1 ).
29.92min (B1).
MS: fi52.4 [M+1]'"
For biological testing, the title compound was transferred into its diacetate
salt, by lyophiliza-
tion from 0.5 M acetic acid (40 ml).
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Example 4
(2E)-5-Amino-5-methylhex-2-enoic acid N-((1R)-1-{N-[(1R)-1-benzyl-2-((2S)-2-
((dimethylamino)methyl)pyrrolidin-1-yl)-2-oxoethyl]-N-methylcarbamoyl}-2-(2-
naphthyl)ethyl)-
5 N-methylamide
/ /
w
H3C CH3 O CH3 O
N
H2N ~ N~ N
CH3 O
N~CH3
I
CH3
(2S)-Pyrrolidine-1,2-dicarboxylic acid 1-tert-butyl ester 2-ethyl ester
~O,~CH3
N ~ '~'I
~ O
O' _O
H C_ I _CH
3 CH3 3
N-tert-Butoxycarbonylpralin (24.38 g, 113 mmol) was dissolved in
dichloromethane (60 ml).
Ethanol (7.9 ml, 135 mmol) and 4-dimethylaminopyridine (1.52 g, 12.5 mmol)
were added.
The solution was cooled to 0 °C. N-(3-dimethylaminopropy!)-N'-
ethylcarbodiimide hydrochlo-
ride (23.88 g, 125 mmol) was added. The reaction mixture was stirred for 16 h,
while it was
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warming up to room temperature. Ethyl acetate (400 ml) was added. It was
washed with a
10% aqueous solution of sodium hydrogen sulfate (300 ml). The aqueous phase
was ex-
tracted with ethyl acetate (3 x 200 ml). The combined organic layers were
washed with a
saturated aqueous solution of sodium hydrogen carbonate (300 ml) and dried
over magne-
slum sulfate. The solvent was removed in vacuo. The crude product was purified
by flash
chromatography on silica (400 g), using ethyl acetate (1:4) as eluent, to give
17.11 g of (2S)-
pyrrolidine-1,2-dicarboxylic acid 1-tert-butyl ester 2-ethyl ester.
'H-NMR (CDCI3): 8 1.28 (m, 3 H); 1.43 and 1.46 (both s, together 9 H}; 2.95
(m, 3 H); 2.22
(m, 1 H); 3.50 (m, 2 H); 4.18 and 4.30 (m and dd, together 3 H).
N-t-Butyloxycarbonyl-(S)-prolinal
,O
N
O- ' O
H3C-~--CH3
CH3
At -78 °C, a 1.2 M solution of diisobutylaluminum hydride(31.7 ml, 38
mmol) in toluene was
added dropwise to a solution of (2S)-pyrrolidine-1,2-dicarboxylic acid 1-tert-
butyl ester 2-
ethyl ester (4.02 g, 16.5 mmol) in diethyl ether (15 ml). The reaction mixture
was stirred for 3
h at -78 °C. Water (9.9 ml) was added dropwise. The reaction mixture
was warmed to room
temperature. The mixture was filtered through a plug of celite. The celite was
washed with
tert-butyl methyl ether (3 x 100 ml). The combined organic layers were dried
over magne-
sium sulfate. The solvent was removed in vacuo, to give 2.34 g of crude N-t-
butyloxy-
carbonyl-(S}-prolinal, which was used for the next step without further
purification.
'H-NMR (CDCI3): 8 1.42 and 1.47 (both s, together 9 H); 1.70 - 2.20 (m, 4 H);
3.20 - 4.30 (m,
3 H); 9.45 and 9.55 (both s, together 1 H).
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{2S)-2-((Dimethylamino)methyl)pyrrolidine-1-carboxylic acid tert-butyl ester
CH3
N~CH3
O' _O
H C_ I _CH
3 CH3 3
Crude N-t-butyloxycarbonyl-(S)-prolinal (2.34 g, 11.7 mmol) was dissolved in
dichloro-
methane (90 ml). A 5.6 M solution of dimethylamine in ethanol (4.19 ml, 23.5
mmol) was
added. 0.4 nm Mol sieves (lO.Og) was added. Sodium triacetoxyborohydride 7.47
g, 35.2
mmol) and glacial acetic acid (1.34 ml, 23.5 mmol) were added successively.
The reaction
mixture was stirred for 3 days. It was filtered through a plug of celite. The
celite was washed
with methanol (150 ml). An 1 N aqueous solution of sodium hydroxide {150 ml)
and tert-butyl
methyl ether (150 ml) were added. The phases were separated. The aqueous phase
was
extracted with tert-butyl methyl ether (3 x 100 ml). The combined organic
layers were dried
over magnesium sulfate. The solvent was removed in vacuo. The crude product
was purified
by flash chromatography on silica (90 g), using dichloromethane/methanol/25%
aqueous
ammonia (100:10:1) as eluent, to give 1.29 g of (2S)-2-
((dimethylamino)methyl)pyrrolidine-1-
carboxylic acid tert-butyl ester.
'H-NMR (CDCI3): b 1.48 (s, 9 H); 1.90 (m, 4 H); 2.15 and 2.23 (AB, 2 H); 2.28
(s, 6 H); 3.31
(br, 2 H); 3.85 (br, 1 H).
N-Dimethyl-N-(((2S)-pyrrolidin-2-yl)methyl)amine
CH3
H~N~CH3
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A 2.7 M solution of hydrogen chloride in ethyl acetate (75 ml, 202 mmol) was
given to a so-
lution of (2S)-2-((dimethylamino)methyl)pyrrolidine-1-carboxylic acid tert-
butyl ester (1.29 g,
5.65 mmol) in ethyl acetate (30 ml). The reaction mixture was stirred for 30
min at room
temperature. The solvent was removed in vacuo to give 1.36 g of the crude
dihydrochoriide
salt of N-dimethyl-N-(((2S)-pyrrolidin-2-yl)methyl)amine, which was used for
the next step
without further purification.
'H-NMR (CDCI3): 8 1.90 (m, 2 H); 2.17 (m, 1 H); 2.40 (m, 1 H); 2.90 (m, 2 H);
3.14 (s, 6 H);
3.55 (m, 2 H); 4.35 (m, 1 H).
N-[(1 R)-1-Benzyl-2-((2S)-2-((dimethylamino)methyl)pyrrolidin-1-yl)-2-
oxoethyi]-N-
methylcarbamic acid tent-butyl ester
CH3 O
H3C~O~N
~N
H3C CH3 O
N~CH3
CH3
At 0 °C, N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride
(1.30 g, 6.76 mmol)
was added to a solution of (2R}-2-(N-{tert-butoxycarbonyl)-N-methylamino)-3-
phenyl-
propionic acid (1.89 g, 6.76 mmol) and 1-hydroxy-7-azabenzotriazole 0.92 g,
6.76 mmol) in
dichloromethane (10 ml). The reaction mixture was stirred for 20 min at 0
°C. A solution of
the crude dihydrochorlide salt of N-dimethyl-N-(((2S)-pyrrolidin-2-
yl)methyl)amine (1.36 g,
6.76 mmol) in dichloromethane (10 ml) and N,N-dimethylformamide (10 ml) and
ethyldiiso-
propylamine (5.75 ml, 33.8 mmol) were added successively. The reaction mixture
was
stirred for 16 h, while it was warming up to room temperature. It was diluted
with ethyl ace-
tate (100 mI) and washed with a saturated aqueous solution of sodium hydrogen
carbonate
{100 ml). The aqueous phase was extracted with ethyl acetate (3 x 80 ml). The
combined
organic layers were dried over magnesium sulfate. The solvent was removed in
vacuo. The
crude product was purified by flash chromatography on silica (90 g), using
dichloro-
methanelmethanol/25% aqueous ammonia (100:10:1 ) as eiuent, to give 2.26 g of
N-[(1 R)-1-
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benzyl-2-((2S)-2-((dimethylamino)methyl)pyrrolidin-1-yl)-2-oxoethylj-N-
methylcarbamic acid
tert-butyl ester.
'H-NMR (CDCI,, selected values): 8 1.20, 1.33, and 1.37 (all s, together 9 H);
2.22 and 2.28
(both s, together 6 H); 2.82 and 2.84 (both s, together 3 H); 4.25 (m, 1 H);
4.80, 5.11, and
5.30 (dd, t, and m, together 1 H); 7.10 - 7.30 (m, 5 H).
C~H~N303 [389.5]
caic. Cfi7.83 H9.06 N10.79
found C67.39 H9.13 N10.73
(2R}-1-{(2S)-2-((Dimethylamino)methyl)pyrrolidin-1-yl)-2-methylamino-3-
phenylpropan-1-one
CH3 O
HN
~N
\ N~CH3
CH3
At 0 °C, trifluoroacetic aad (8 ml) was added to a solution of N-((1 R)-
1-benzyl-2-((2S}-2-
((dimethylamino)methyl)pyrrolidin-1-y1~2-oxoethyl]-N-methylcarbamic acid tert-
butyl ester
(2.26 g, 5.80 mmol) in dichloromethane (8 ml).~The reaction mixture was
stirred for 20 min at
0 °C. The solvent was removed in vacuo. Dichloromethane (70 ml) was
added, and the sol-
vent was removed in vacuo. The latter procedure was repeated two times. The
crude prod-
uct was purfied by flash chromatography on silica (90 g), using
dichloromethanelmetha-
noll25°~ aqueous ammonia (100:10:1) as eiuent, to give 1.24 g of (2R)-1-
((2S)-2-((dimethyl-
amino)methyl)pyrrolidin-1-yl)-2-methylamino-3-phenylpropan-1-one.
'H-NMR (CDC13, selected values): 8 2.33 (s, 3 H); 2.43 (s, 6 H); 3.25 (m, 3
H); 4.17 (m, 1 H);
7.25 (m, 5 H).
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N-((1 R)-1-{N-[(1 R)-1-Benzyl-2-((2S)-2-((dimethylamino)methyl)pyrrolidin-1-
yl)-2-oxoethyf]-N-
methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-methylcarbamic acid tert-butyl ester
H C CH3 O CH3 O
H3C_ _O_ -N~ N N
l ll
CH3 O
\ N~CH3
I
CH3
At 0 °C, N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride
(530 mg, 2.76 mmol)
was added to a solution of (2R)-2-(N-(tert-butoxycarbonyl)-N-methylamino)-3-(2-
naphthyl)
10 propionic acid (911 mg, 2.76 mmol) and 1-hydroxy-7-azabenzotriazole (376
mg, 2.76 mmol)
in dichloromethane (5 ml). The reaction mixture was stirred for 20 min at 0
°C. A solution of
(2R)-1-((2S)-2-((dimethylamino)methyl)pyrrolidin-1-yl)-2-methyiamino-3-
phenylpropan-1-one
(800 mg, 2.76 mmol) in dichioromethane (5 ml) and N, N-dimethylformamide (5
mi) and eth-
yldiisopropylamine (0.71 ml, 4.15 mmol) were added successively. The reaction
mixture was
15 stirred for 3 days, white it was warming up to room temperature. It was
diluted with ethyl
acetate (100 ml) and washed with a saturated aqueous solution of sodium
hydrogen carbon-
ate (100 ml). The aqueous solution was extracted with ethyl acetate (3 x 70
ml). The com-
bined organic layers were dried over magnesium sulfate. The solvent was
removed in vacuo.
The crude product was purified by flash chromatography on silica (90 g), using
dichloro-
20 methanelmethanol/25% aqueous ammonia (200:10:1 ) as eluent, to give 1.37 g
of N-((1 R)-1-
{N-[( 1 R)-1-benzyl-2-((2S)-2-({dimethylamino)methyl)pyrrolidin-1-yl)-2-
oxoethyl]-N-methyl-
carbamoyl}-2-(2-naphthyl)ethyl)-N-methylcarbamic acid tert-butyl ester.
'H-NMR (CDCI3, selected values): 8 0.64 (m, 1 H); 1.10, 1.29, 1.36, and 1.47
(all s, together
25 9 H); 4.99, 5.09, 5.45, and 5.53 (t, t, m, and t, together 2 H); 7.10 -
7.90 (m, 12 H).
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(2R)-N-[(1 R)-1-Benzyl-2-((2S)-2-((dimethyiamino)methyl)pyrrolidin-1-yl)-2-
oxoethyl]-N-
methyl-2-(methylamino)-3-(2-naphthyl)propionamide
CH3 O
N
HN~ N
CH3 'OI
\ N~CHs
CH3
At 0 °C, trifluoroacetic acid (10 ml) was added to a solution of N-
((1R)-1-{N-[(1R)-1-benzyl-2-
((2S)-2-((dimethylamino)methyl)pyrrolidin-1-yl)-2-oxoethyl]-N-methylcarbamoyl}-
2-(2-
naphthyl)ethyl)-N-methyicarbamic acid tert-butyl ester (1.37 g, 2.28 mmol) in
dichloro-
methane (10 ml). The reaction mixture was stirred fo 75 min at 0 °C.
The solvent was re-
moved in vacuo. The residue was dissolved in dichloromethane (70 ml) and the
solvent was
removed in vacuo. The latter procedure was repeated two times. The crude
product was pu-
rifled by flash chromatography on silica (90 g), using
dichloromethane/methanoll25% aque-
ous ammonia (100:10:1) as eluent, to give 692 mg of (2R)-N-[(1R)-1-benzyl-2-
((2S)-2-
((dimethylamino)methyl)pyrrolidin-1-y1~2-oxoethyl]-N-methyl-2-(methylamino)-3-
(2-
naphthyl)propionamide.
'H-NMR (CDCl3, selected values): 8 1.85 and 2.01 (both s, together 3 H); 2.20
and 2.31
(both s, together 6 H); 3.65 and 3.80 (both t, 1 H); 4.04 and 4.45 (both m,
together 1 H); 5.60
and 5.91 (t and dd, together 1 H); 7.10 - 7.90 (m, 12 H).
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{(3E)-4-[N-((1 R)-1-{N-[(1 R}-1-Benzyl-2-((2S)-2-
((dimethytamino)methyl)pyrrolidin-1-yi)-2
oxoethyl]-N-methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-methylcarbamoyl]-1,1-
dimethylbut-3
enyl}carbamic acid tert-butyl ester
H C CH3 O H3C CH3 O CH3 O
H C~O~N ~ N~ N N
I II
CH3 O
/ \ N~CH3
CH3
At 0°C, N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride
(132 mg, 0.69 mmol)
was added to a solution of (2E)-5-(tert-butoxycarbonylamino)-5-methylhex-2-
enoic acid (168
mg, 0.69 mmol) and 1-hydroxy-7-azabenzotriazoie (94 mg, 0.69 mmol) in
dichloromethane
(5 ml). The reaction mixture was stirred for 20 min at 0 °C. A solution
of (2R)-N-[(1 R)-1-
benzyl-2-((2S)-2-((dimethylamino)methyl)pyrrofidin-1-yl)-2-oxoethyl]-N-methyl-
2-(methyl-
amino)-3-(2-naphthyl)propionamide (345 mg, 0.69 mmol) in dichioromethane (10
ml) and
N,N-dimethylformamide (5 ml) and ethyldiisopropylamine were added
successively. The re-
action mixture was stirred for 16 h, while it was warming up to room
temperature. It was di-
luted with ethyl acetate (70 ml) and washed with a saturated aqueous solution
of sodium hy-
drogen carbonate (70 ml). The aqueous phase was extracted with ethyl acetate
(3 x 50 ml).
The combined organic layers were dried over magnesium sulfate. The solvent was
removed
in vacuo. The crude product was purified by flash chromatography on silica (40
g), using di-
chloromethanelmethanoU25°~ aqueous ammonia (100:10:1 ) as eiuent, to
give 491 mg of
{(3E)-4-[N-((1 R)-1-{N-[(1 R)-1-benzyl-2-((2S)-2-
((dimethylamino)methyl)pyrrolidin-1-yl)-2-
oxoethyl]-N-methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-methylcarbamoyl]-1,1-
dimethylbut-3-
enyl}carbamic acid tert-butyl ester.
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'H-NMR (CDC13, selected values): b 1.30 and 1.32 (both s, together 6 H); 1.45
(s, 9 H); 1.60
(m, 2 H); 4.00 (m, 1 H); 4.48 (m, 1 H); 5.48 (dd, 1 H); 5.92 (dd, 1 H); 6.11
and 6.20 (both d,
together 1 H); 6.82 and 6.92 (both m, together 1 H); 7.10 - 7.90 (m, 12 H).
At 0 °C, trifluoroacetic acid (7 ml) was added to a solution of {(3E)-4-
[N-((1R)-1-{N-[(1R)-1-
benzyl-2-((2S)-2-((dimethylamino)methyl)pyrrolidin-1-yl)-2-oxoethyl]-N-
methyicarbamoyl}-2-
(2-naphthyl)ethyl)-N-methylcarbamoyl]-1,1-dimethylbut-3-enyl}carbamic acid
tert-butyl ester
(491 mg, D.68 mmol) in dichloromethane (7 ml). The reaction mixture was
stirred for 60 min
at 0 °C. The solvent was removed in vacuo. The residue was dissolved in
dichloromethane
(100 ml) and the solvent was removed in vacuo. The latter procedure was
repeated two
times. The crude product was purified by flash chromatography on silica (40
g), using di-
chloromethane/methanol/25% ammonia (100:10:1) as eluent, to give 285 mg of the
title
compound.
'H-NMR (CDCI3, selected values): 8 0.55 {m, 1 H); 1.11, 1.12, and 1.17 (all s,
together 6 H);
2.25 (s, 6 H); 2.45 (s, 3 H); 2.85 (s, 3 H); 4.02 (m, 1 H); 5.48 (dd, 1 H);
5.80 and 5.93 (m,
and dd, together 1 H); 6. i 0 and 6.18 (both d, together 1 H); 6.87 and 7.OD
(both m, together
1 H); 7.10 - 7.90 (m, 12 H).
HPLC 27.97 min (A1 ).
27.80 min (B1).
MS: 626.4 [M+1 ]'
For biological testing, the title compound was transferred into its diacetate
salt by iyophifiza-
tion from 0.5 acetic acid (40 ml).
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Example 5
N-((1 R)-1-{N-[(1 R)-1-Benzyl-2-((2S}-2-((dimethylamino)methyl)pyrrolidin-1-
yl)-2-oxoethyl]-N-
methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-methyl-3-
((methylamino)methyl)benzamide
/ /
O CH3 O
HN ~ N~ N N
I ~ I Il
CH3 / CH3 O
N~CH3
I
CH3
N-~3-[N-((1R)-1-{N-[(1R)-1-Benzyl-2-((2S)-2-((dimethylamino)methyl)pyrrolidin-
1-yl)-2-
oxoethyl]-N-methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-methylcarbamoyl]benzyl}-N-
methyl-
carbamic acid tert-butyl ester
/ /
H C CH3 O O CH3 O
H3CI _O- _N ~ N~ N N
I I I II
CH3 / CH3 O
N~CH3
CH3
At 0 °C, N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride
(132 mg, 0.69 mmol)
was added to a solution of 3-(N-(tert-butoxycarbonyl)-N-methylamino}benzoic
acid (183 mg,
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0.69 mmol) and 1-hydroxy-7-azabenzotriazole (94 mg, 0.69 mmol) in
dichloromethane (5
ml). The reaction mixture was stirred for 20 min at 0 °C. A solution of
(2R)-N-[(1R)-1-benzyl-
2-((2S)-2-((dimethylamino)methyl)pyrrolidin-1-yl)-2-oxoethylj-N-methyl-2-
(methylamino)-3-(2-
naphthyl)propionamide (345 mg, 0.69 mmof) in dichloromethane (10 ml) and N,N-
dimethyl-
5 formamide (5 ml) and ethyldiisopropylamine (0.118 ml) were added
successively. The reac-
tion mixture was stirred for 16 h, while it was warming up to room
temperature. It was diluted
with ethyl acetate and washed with a saturated aqueous solution of sodium
hydrogen car-
bonate (70 ml). The aqueous phase was extracted with ethyl acetate (3 x 50
ml). The com-
bined organic layers were dried over magnesium sulfate. The solvent was
removed in vacuo.
10 The crude product was purified by flash chromatography on silica (40 g),
using dichloro-
methane/methanoll25% aqueous ammonia (100:10:1) as eluent, to give 524 mg of N-
{3-[N-
((1 R)-1-{N-[(1 R)-1-benzyl-2-((2S)-2-((dimethylamino)methyl)pyrrolidin-1-yl)-
2-oxoethylj-N-
methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-methylcarbamoyljbenzyl}-N-
methylcarbamic acid
tert-butyl ester.
'H-NMR (CDC13, selected values): 8 0.72 (m, 1 H); 1.45 (br, 9 H); 3.18 (br, 6
H); 4.05 (m, 1
H); 4.32 and 4.40 (both br, together 2 H); 5.60 (dd, 1 H); 5.95 (m, 1 H); 6.80
- 6.90 (m, 16
H).
At 0 °C, trifluoroacetic acid (7 ml) was added to a solution of N-{3-[N-
((1R)-1-{N-[(1R)-1-
benzyl-2-((2S)-2-((dimethylamino)methyl)pyrrolidin-1-yl)-2-oxoethyl]-N-
methylcarbamoyl}-2-
(2-naphthyl)ethyl}-N-methylcarbamoyl]benzyl}-N-methylcarbamic acid tert-butyl
ester (523
mg, 0.70 mmol) in dichloromethane (7 ml). The reaction mixture was stirred for
25 min at 0
°C. The solvent was removed in vacuo. The residue was dissolved in
dichloromethane (90
ml), and the solvent was removed in vacuo. The latter procedure was repeated
two times.
The crude product was purified by flash chromatography on silica (40 g), using
dichloro-
methane/methanol/25% aqueous ammonia (100:10:1) as eluent, to give 436 mg of
the title
compound.
'H-NMR (CDCI3, selected values): 8 0.87 (m, 1 H); 1.22 (m, 1 H); 1.45 (m, 1
H); 1.67 (m, 1 );
4.09 (m, 1 H); 5.53 and 5.90 (dd and m, together 2 H); 6.80 - 7.90 (m, 16 H).
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HPLC 28.43 min (A1 ).
30.63 min (B1).
MS: 648.4 [M+1]'
For biological testing, the title compound was transferred into its diacetate
salt by lyophiliza-
tion from 0.5 M acetic acid (40 ml).
Example 6
(2E)-5-Amino-5-methylhex-2-enoic acid N-((1 R)-1-{N-[(1 R)-1-benzyl-2-(4-
(dimethyl-
amino)piperidin-1-yl)-2-oxoethyl]-N-methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-
methylamide.
H3C CH3 ~ CH3 O
H2N \ N N
l~ N
CH3 p N~CH3
/ CH3
The title compound was prepared as in example 1 using 4-
(dimethylamino)piperidine hydro
chiorid salt, (2R)-2-(N-(tent-butoxycarbonyl)-N-methylamino)-3-phenylpropionic
acid, (2R)-2-
(N-(tert-butoxycarbonyl)-N-methylamino-3-(2-naphthyl))propionic acid, and (2E)-
5-
(butoxycarbonylamino)-5-methylhex-2-enoic acid.
'H-NMR : (CDCI,; selected values) b 1.40 (s, 6 H); 2.00 (s, 6 H); 4.42 - 4,85
(2 H); 5.45 -
5.90 (m, 2 H); 6.28 (dd, 1 H); 6.85(m, 1 H); 7.10 - 7,85(m, 12 H)
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MS(ES) : mlz 626.2 (M+H)'
Example 7
(2E)-5-Amino-5-methylhex-2-enoic acid N-methyl-N-[(1 R)-1-(N-methyl-N-{(1 R)-1-
[N-methyl-
N-(1-methylpiperidin-4-yl)carbamoyl]-2-phenylethyl}carbamoyl)-2-(2-
naphthyl)ethyl]amide
H3C CH3 ~ CH3 O N~CH3
H2N \ N~ N N
I II I
CH3 ~ CH
The title compound was prepared as in example 1 using 1-methyl-4-
(methylamino)piperidine,
(2R)-2-(N-(tert-butoxycarbonyl)-N-methylamino)-3-phenylpropionic acid, (2R)-2-
(N-(tert
butoxycarbonyl)-N-methylamino-3-(2-naphthyl))propionic acid, and (2E)-5-
(butoxycarbonyl-
amino)-5-methylhex-2-enoic acid.
'H-NMR (CDCI3; selected values) 8 5.50 - 6.08 (m, 2 H); 6.20 - 6.70 {m, 2 H);
7.10 - 7.85 (m,
12 H)
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Example 8
3-Aminomethyl-N-(( 1 R)-1-{N-[( 1 R)-1-benzyl-2-(4-methylpiperazin-1-yl)-2-
oxoethyl]-N-
methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-methylbenzamide
O CH3 O
H2N ~ N N N
W
CH3 O ~N~CH
3
The title compound was prepared as in example 1 using N-methylpiperazine, (2R)-
2-(N-(tert-
butoxycarbonyl)-N-methylamino)-3-phenylpropionic acid, (2R)-2-(N-(tert-
butoxycarbonyl)-N-
methylamino-3-(2-naphthyl))propionic acid, and 3-((tert-
butoxycarbonylamino)methyl)benzoic
acid.
'H-NMR (CDC13; selected values) 8 3.30 (m, 1 H); 3.50 (dd, 1 H); 3.75 (m, 1
H); 3.95 (s, 2
H); 5.78 (t, 1 H); 3.88 (m, 1 H); 7.00 - 7.80 (16 H).
HPLC : 24.55 min (A1 ).
26.52 min (B1).
MS(ES) : m/z = 606.4 [M+H]'.
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Example 9
(2E)-5-Amino-5-methylhex-2-enoic acid N-((1 R)-1-{N-[(1 R)-1-benzyi-2-(4-
methyipiperazin-1-
yl)-2-oxoethyl]-N-methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-methylamide
H3C CH3 O CH3 O
H2N ~ N~ N N
I II
CH3 O ~N~CH
3
The title compound was prepared as in example 1 using N-methylpiperazine, (2R}-
2-(N-(tert-
butoxycarbonyl)-N-methylamino)-3-phenylpropionic acid, (2R)-2-(N-(tert-
butoxycarbonyl)-N-
methy!amino-3-(2-naphthyl))propionic acid, and (2E)-5-(butoxycarbony!amino)-5-
methylhex-
2-enoic acid.
'H-NMR (CDCI3; selected values) s 1.24 (s, 6 H); 1.65 (s, 3 H); 2.35 (s, 3 H);
2.80 (s, 3 H);
5.68 (dd, 1 H); 5.78 (dd, 1 H); 6.18 (dd, 1 H); 6.95(m, 1 H); 7.15 - 7.80 (m,
12 H).
HPLC : 25.03 min (A1 ).
27.50 min (B1 ).
MS(ES) : mlz = 598.4 [M+H]'.
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Example 10
(2E}-5-Amino-5-methylhex-2-enoicacid N-methyl-N-((1R)-1-{N-methyl-N-j(1 R)-2-
phenyl-1-
((2,2,6,6-tetramethylpiperidin-4-yl)carbamoyl)ethyl]carbamoyl}-2-(2-
naphthyl)ethyl)amide
H3C CH3
H C CH O CH3 O ~NH
3 3 N CH3
HZN ~ N~ H CH
CH3 O s
The title compound was prepared as in example 1 using 4-amino-2,2,6,6-
tetramethyl-
piperidin, (2R)-2-(N-(tent-butoxycarbonyl)-N-methylamino)-3-phenyipropionic
acid, (2R)-2-(N-
(tert-butoxycarbonyl)-N-methylamino-3-(2-naphthyl))propionic acid, and (2E)-5-
(butoxy-
carbonylamino)-5-methylhex-2-enoic acid.
'H-NMR (CDCI3; selected values) 8 1.25 (s, 6 H); 1.40 (two s, 6 H); 1.52 (two
s, 6 H); 2.92
(s, 3 H); 3.02 (two s, 3 H); 5.10 (dd, 1 H); 5.50 (dd, 1 H); 6.15 (d, 1 H);
6.75 (m, 1 H); 7.00
15 8.00 (m, 12 H).
HPLC : 29.27min (A1 }.
31.67min (B1).
MS(ES) : m/z = 654.8 jM+H)'.
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Example 11
3-Aminomethyl-N-methyl-N-({1 R)1-{N-methyl-N-[(1 R)-2-phenyl-1-((2,2,6,6-
tetramethylpiperidin-4-yl)carbamoyl)ethyl]carbamoyl}-2-(2-
naphthyl)ethyl)benzamide
O CH3 O H3C CH
3
w HZN ~ N N N NH
I '~ H
/ CH3 O CH3
H3C
The title compound was prepared as in example 1 using 4-amino-2,2,6,6-
tetramethyl-
piperidin, (2R)-2-(N-(tert-butoxycarbonyl)-N-methylamino)-3-phenylpropionic
acid, (2R)-2-(N-
(tert-butoxycarbonyl)-N-methylamino-3-(2-naphthyl))propionic acid, and 3-
((tert-butoxy-
carbonylamino)methyl)benzoic acid.
'H-NMR (CDC13; selected values) S 3.60 - 3.85 (m, 2 H); 3.90 - 4.30 (m, 1 H);
5.25 -5.95 (m,
2 H); 6.70 - 7.90 (m, 16 H).
HPLC : 29.27 min (A1 ).
31.55 min (B1 ).
MS(ES) : mlz = 662.4 [M+H]'.
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Example 12
(2E)-5-Amino-3,5-dimethylhex-2-enoic acid N-methyl-N-((1 R)-1-{N-methyl-N-[(1
R}-2-phenyl
1-((2,2,6,6-tetramethylpiperidin-4-yl)carbamoyl)ethyljcarbamoyl}-2-(2-
naphthyl)ethyl)amide
i
i
H3C CH CHs O CH3 O H3C CH3
v
H2N \ N~ N N NH
I II H
CH3 O CH3
H3C
The title compound was prepared as in example 1 using 4-amino-2,2,6,6-
tetramethyl-
piperidin, (2R)-2-(N-(tert-butoxycarbonyl)-N-methylamino}-3-phenylpropionic
acid, (2R)-2-(N-
(tert-butoxycarbonyl)-N-methylamino-3-(2-naphthyl))propionic acid, and (2E)-5-
(butoxy-
carbonylamino)-3,5-methylhex-2-enoic acid.
'H-NMR (CDCI3; selected values) 8 3.92 - 4.30 (m, 1 H); 5.05 - 5.88 (m, 3H);
7.00 - 7.80 (m,
12 H}.
HPLC : 29.80 min {A1 ).
32.43 min (B1 ).
MS(ES) : m/z = 668.4 [M+H]'.
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Example 13
(2E)-4-(1-Arninocyclobutyl)but-2-enoic acid N-((1R)1-{N-[(1R)1-benzyl-2-(4-
methylpiperazin-
1-yl)-2-oxoethylJ-N-methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-rnethylamide
.
O CH3 O
N
w... H2N ~ N~ N
CH3 O ~N~CH
3
The title compound was prepared as in example 1 using N-methyipiperazine, (2R)-
2-(N-(tert-
butoxycarbonyl)-N-methylamino)-3-phenylpropionic acid, (2R)-2-(N-(tert-
butoxycarbonyl)-N-
methylamino-3-(2-naphthyl))propionic acid, and (2E)-4-(1-
(butoxycarbonylamino)cyclo-
butyl)but-2-enoic acid.
'H-NMR (CDCI3; selected peaks) 8 1.62 (s, 3 H); 2.35 (s, 3 H); 2.80 (s, 3 H);
5.70 (dd, 1 H);
5.80 (dd, 1 H); 6.22 (d, 1 H); 6.98 (m, 1 H); 7.15 - 7.80 (m, 12 H).
HPLC : 25.88 min (A1 ).
28.65 min (B1 ).
MS(ES) : m/z = 610.4 [M+H]'.
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Example 14
(2E)-5-Amino-3,5-dimethylhex-2-enoic acid N-((1R)1-{N-[(1R)1-benzyl-2-(4-
methylpiperazin-
1-yl)-2-oxoethyl)-N-methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-methylamide
H C CH CHs O CH3 O
3 3 I
H2N \ N~ N N
I II
CH3 O ~N~CH
3
The title compound was prepared as in example 1 using N-methylpiperazine, (2R)-
2-(N-{tert-
butoxycarbonyl)-N-methylamino)-3-phenyipropionic acid, (2R)-2-(N-(ter#-
butoxycarbonyl)-N-
methy!amino-3-(2-naphthyl))propionic acid, and (2E)-5-{butoxycarbony!amino)-
3,5-methyl-
hex-2-enoic acid.
'H-NMR (CDCI3; selected values) b 1.18 (s, 6 H); 1.68 (s, 3 H); 1.95 (s, 3 H);
2.30 (s, 3 H);
2.85 (s, 3 H); 3.40 (dd, 1 H); 3.54 - 3.75 (m, 2 H); 5.68 - 5.85 (m, 3 H);
7.15 - 7.80 (m, 12 H).
HPLC : 25.70 min (A1 ).
28.27 min (B1 ).
MS(ES) : mlz = 612.4 [M+H)'.
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Example 15
(2E)-4-(1-Aminocyctobutyl)but-2-enoic acid N-((1 R)-1-{N-[(1 R)-1-benzyl-2-{4-
hydroxypiperidin-1-yl)-2-oxoethyl]-N-methylcarbamoyl}-2-(biphenyl-4-yl)ethyl)-
N-
methylamide
/ \
/ \
O CH3 O
H2N ~ N ~ N N
CH3 O OH
\ /
This compound was prepared as in example 1 but using 4-hydroxypiperidine, (2R)-
{N-tert-
butoxycarbonyl-N-methylamino)-3-phenyipropionic acid and (2R~2-(N-tert-
butoxycarbonyl-
~0 N-methylamino)-3-(biphenyl-4-yl)propionic acid and (2E)-4-{1-(tert-
butoxycarbonylamino)cyclobutyl)but-2-enoic acid as starting materials.
ESMS: 637. 4 (M+H)'
HPLC: r, = 33.58 min. (A1 )
15 HPLC: r, = 34.95 min. (B1 )
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Example 16
(2E}-5-Amino-5-methylhex-2-enoic acid N-({1 R)-1-{N-[(1 R}-1-benzyl-2-(4-
hydroxypiperidin-1-
yl)-2-oxoethyl]-N-methylcarbamoyl}-2-(biphenyl-4-yl)ethyl)-N-methylamide
/ \
H3C CH3 O CH3 O
H2N ~ N~N N
CH3 O
OH
\ /
This compound was prepared as in example 1 but using 4-hydroxypiperidine, (2R)-
2-{N-tert-
butoxycarbonyl-N-methylamino)-3-phenylpropionic acid and (2R)-2-{N-tert-
butoxycarbonyl-
N-methylamino)-3-(biphenyl-4-yl)propionic acid and (2E)-5-tert-
Butoxycarbonylamino-5-
methyfhex-2-enoic acid as starting materials
ESMS: 625.4 (M+H)'
HPLC: r, = 32.65 min. (A1 )
HPLC: r, = 34.02 min. (B1)
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Example 17
(2E)-5-Amino-3,5-dimethylhex-2-enoic acid N-((1R)-1-{N-[(1R)-1-benzyl-2-(4-
hydroxy-
piperidin-1-yl)-2-oxoethylj-N-methylcarbamoyl}-2-(biphenyl-4-yl)ethyl)-N-
methylamide
.,.. H3C CHCHa O CH3 O
H2N \ N~N N
CH3 O OH
\ /
This compound was prepared as in example 1 but using 4-hydroxypiperidine, (2R)-
2-(N-tert-
butoxycarbonyl-N-methylamino)-3-phenylpropionic acid and (2R)-2-(N-tert-
butoxycarbonyl-
N-methyiamino)-3-(biphenyl-4-yl)propionic acid and (2E)-5-tert-
butoxycarbonylamino-3,5-
dimethylhex-2-enoic acid as starting materials.
ESMS: 639.4 (M+H)'
HPLC: r, = 33.29 min. (A1 )
HPLC: r, = 36.40 min. (B1 )
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Example 18
(2E)-5-Amino-5-methylhex-2-enoic acid N-((1R)-1-{N-[(1R)-1-benzyl-2-(4-
hydroxypiperidin-1-
yl)-2-oxoethyl]-N-methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-methylamide
H3C CH3 O CH3 O
H2N ~ N~N N
i
CH3 O OH
This compound was prepared as in example 1 but using 4-hydroxypiperidine, (2R)-
2-(N-tert-
butoxycarbonyl-N-methylamino)-3-phenylpropionic acid and (2R)-2-(N-tert-
butoxycarbonyl-
N-methylamino)-3-(2-naphthyl)propionic acid and (2E)-5-tert-
Butoxycarbonylamino-5-methyl-
hex-2-enoic acid as starting materials.
ESMS: 599.4 (M+H)'
HPLC: r, = 29.88 min. (A1 )
'f 5
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Example 19
(2E)-5-Amino-3,5-dimethylhex-2-enoic acid N-((1R}-1-{N-((1R)-1-benzyl-2-(4-
hydroxypiperidin-1-yl)-2-oxoethyl}-N-methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-
methylamide
H C CH CHs O _ CH3 O
HN 3\ N N N
I
CH3 O OH
This compound was prepared as in example 1 but using 4-hydroxypiperidine, (2R)-
2-(N-tert-
butoxycarbonyl-N-methyfamino)-3-phenylpropionic acid and (2R)-2-(N-tert-
butoxycarbonyl-
N-methylamino)-3-(2-naphthyl)propionic acid and (2E)-5-tert-
Butoxycarbonylamino-3,5-
dimethylhex-2-enoic acid as starting materials.
ESMS:613.4 (M+H)'
HPLC: r, = 30.58 min. (A1 )
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Example 20
(2E)-4-(1-Aminocyclobuty!)but-2-enoic acid N-((1 R)-1-{N-[(1 R}-1-benzyl-2-(4-
hydroxy-
piperidin-1-yl)-2-oxoethyl]-N-methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-
methyiamide
O CH3 O
H2N ~ N~N N
CH3 O OH
This compound was prepared as in example 1 but using 4-hydroxypiperidine, (2R)-
2-(N-tert-
butoxycarbonyl-N-methylamino)-3-phenyipropionic acid and (2R)-2-(N-tert-
butoxycarbonyl-
N-methylamino)-3-(2-naphthyl)propionic acid and (2E)-4-(1-(tert-
butoxycarbonylamino)cyclo-
butyl)but-2-enoic acid.
ESMS: 611.4 (M+H)'
HPLC: r, = 30.82 min. (A1 )
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Example 21
(2E)-5-Amino-5-methylhex-2-enoic acid N-((1 R)-1-{N-[(1 R)-1-(4-fluorobenzyl)-
2-(4-hydroxy-
piperidin-1-yl)-2-oxoethyl]-N-methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-
methylamide
/ / 1
H3C CH3 O CH3 O
H2N ~ N~N N
CH3 O OH
1
F
This compound was prepared as in example 1 but using 4-hydroxypiperidine, (2R)-
2-(N-tert-
butoxycarbonyl-N-methylamino)-3-(4-fluorophenyl)propionic acid and (2R)-2-(N-
tert-butoxy-
carbonyl-N-methylamino)-3-(2-naphthyl)propionic acid and (2E)-5-tert-
butoxycarbonylamino-
5-methylhex-2-enoic acid as starting materials.
ESMS: 617.4 (M+H)'
HPLC: r, = 30.27 min. (A1)
HPLC: r, = 31.60 min. (B1)
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Example 22
(2E)-5-Amino-3,5-dimethylhex-2-enoic acid N-((1 R)-1-{N-[(1 R)-1-(4-
fluorobenzyl)-2-(4-
hydroxypiperidin-1-yl)-2-oxoethyl]-N-methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-
methyfamide
H3C CH CH3 O CH3 O
3 I
H2N ~ N~N N
CH3 O
OH
F
This compound was prepared as in example 1 but using 4-hydroxypiperidine, (2R)-
2-(N-tert-
butoxycarbonyl-N-methylamino)-3-(4-fluorophenyl)propionic acid and (2R}-2-(N-
tert-butoxy-
carbonyl-N-methylamino)-3-(2-naphthyl)propionic acid and (2E)-5-tert-
butoxycarbonylamino-
3,5-dimethylhex-2-enoic acid as starting materials.
ESMS: 631.4 (M+H)'
HPLC: r, = 30.98 min. (A1 )
HPLC: r, = 32.38 min. (B1)
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Example 23
(2E)4-(1-Aminocyclobutyl)but-2-enoic acid N-((1R)-1-{N-[(1R)-1-benzyl-2-(4-
(dimethyl-
amino)piperidin-1-yl)-2-oxoethyl]-N-methylcarbamoyl}~2-(2-naphthyl)ethyl)-N-
methylamide
O CH3 O
H2N ~ N N N
~..Y. CH3 O N.CH3
CH3
The title compound was prepared as in example 1 using 4-N,N-
dimethylpiperazine, (2R)-2-
(N-(tert-butoxycarbonyl)-N-methylamino)-3-phenylpropionic acid, (2R)-2-(N-
(tert-butoxy-
carbonyl)-N-methylamino-3-(2-naphthyl))propionic acid, and (2E)-4-(1-
(butoxycarbonyl-
amino)cyclobutyl)but-2-enoic acid.
1 H-NMR (CDCI3; selected peaks) d 1.90 (s, 3 H); 2.38 (s, 3 H); 2.45 and 2.47
(two s, 3 H)
2.78 and 2.80 (two s, 3 H); 6.32 (dd, 1 H); 6.90 (m, 1 H); 7.15 - 7.84 (m, 12
H).
HPLC : 26.72 min (A1 ).
MS(ES) : m/z = 638.4 [M+H]+.
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Example 24
(2E)-5-Amino-5-methylhex-2-enoic acid N-((1 R)-1-{N-[(2R)-2-(4-
hydroxypiperidin-1-yl)-2-oxo-
1-((2-thienyl)methyl)ethyl)-N-methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-
methylamide
H3C CH3 O CH3 O OH
H2N ~ N II N N
CH3 O
'J
This compound was prepared as in example 1 but using 4-hydroxypiperidine, (2R)-
2-(N-tert-
butoxycarbonyl-N-methylamino)-3-(2-thienyl)propionic acid and (2R)-2-(N-tert-
butoxy-
carbonyl-N-methyfamino)-3-(2-naphthyl)propionic acid and (2E)-5-tert-
butoxycarbonyiamino-
5-methylhex-2-enoic acid as starting materials.
ESMS: 605.4 (M+H)'
HPLC: rt = 29.07 min. (A1 )
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Example 25
(2E)-5-Amino-3,5-dimethylhex-2-enoic acid N-((1R)-1-(N-[(2R)-2-(4-
hydroxypiperidin-1-yl)-2-
oxo-1-((2-thienyl)methyl)ethyl)-N-methylcarbamoyl}-2-(2-naphthyl)ethyl)-N-
methylamide
k
H C CH3CH3 O CH3 O OH
3
H2N \ N~N N
CH3 O
S
This compound was prepared as in example 1 but using 4-hydroxypiperidine, (2R)-
2-(N-tert-
butoxycarbonyi-N-methylamino)-3-(2-thienyl)propionic acid and (2R)-2-(N-tert-
butoxy-
carbonyl-N-methylamino)-3-(2-naphthyl)propionic acid and (2E)-5-tert-
butoxycarbonylamino-
3,5-dimethylhex-2-enoic acid as starting materials.
ESMS: 619.4 (M+H)+
HPLC: r, = 29.76 min. (A1 )
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Example 26
(2E)-5-Amino-5-methylhex-2-enoic acid N-((1R)-2-(biphenyl-4-yl)-1-{N-[(2R)-2-
(4-
hydroxypiperidin-1-yl)-2-oxo-1-((2-thienyl)methyl)ethyl]-N-
methylcarbamoyl}ethyl}-N-
methylamide
b
H C CH O r CH3 O OH
3 3 I
H2N ~ N~N N
I
CH3 O
This compound was prepared as in example 1 but using 4-hydroxypiperidine, (2R)-
2-(N-tert-
butoxycarbonyl-N-methylamino)-3-(2-thienyl)propionic acid and (2R)-2-(N-tert-
butoxy-
carbonyl-N-methylamino)-3-(biphenyl-4-yl)propionic acid and (2E)-5-tert-
butoxycarbonyl-
amino-5-methyihex-2-enoic acid as starting materials.
ESMS: 631.2 (M+H)'
HPLC: r, = 32.20 min. (A1 )
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Example 27
(2E)-5-Amino-3,5-dimethylhex-2-enoic acid N-((1 R)-2-(biphenyl-4-yl)-1-{N-[(1
R)-2-(4-
hydroxypiperidin-1-yl)-2-oxo-1-((2-thienyl)methyl)ethylj-N-
methylcarbamoyl}ethyl)-N-
methylamide
H3C CH3 CH3 O CH3 O
HZN \ N~N N
CH3 O
OH
S /
This compound was prepared as in example 1 but using 4-hydroxypiperidine, (2R)-
2-(N-tert-
butoxycarbonyl-N-rnethylamino)-3-{2-thienyl)propionic acid and (2R)-2-(N-tert-
butoxy-
carbonyl-N-methylamino)-3-(biphenyl-4-yl)propionic acid and {2E)-5-tert-
butoxycarbonyl-
amino-3,5-dimethylhex-2-enoic acid as starting materials.
ESMS: 465.4 (M+H)'
HPLC: r, = 32.89 min. (A1 )
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Example 28
(2E)-5-Methyl-5-(methylamino)hex-2-enoic acid N-((1R)-1-{N-[(1R)-1-benzyl-2-(4-
hydroxypiperidin-1-yl)-2-oxoethyl]-N-methylcarbamoyl}-2-(biphenyl-4-yl)ethyl)-
N-
methylamide
i
i
CH3 O CHs O
~N
HCH CH3 \ CH O N. OH
3 3
The title compound was prepared as in example 1 but using 4-hydroxypiperidine,
(2R)-2-(N-
tert-butoxycarbonyl-N-methylamino)-3-phenylpropionic acid and (2R}-2-(N-tert-
butoxy-
carbonyl-N-methylamino)-3-(biphenyl-4-yl)propionic acid and (2E)-5-(N-(tert
Butoxycar-
bonyl)-N-methylamino)-5-methylhex-2-enoic acid as starting materials
MS: m/z: fi39.4 (M+H)'
HPLC: Method A1:R, = 32.94 min
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Example 29
(2E)-4-( i -Aminocyclobutyl)but-2-enoic acid (( 1 R)-1-{N-[( 1 R)-1-benzyl-2-
(4-hydroxypiperidin-
1-yl)-2-oxoethylJ-N-methylcarbamoyl}-2-(biphenyl-4-yl)ethyl)amide
/ \
/ \
O CH3 O
H2N ~ N ~ N N
H p
OH
\ /
HPLC: Rt = 31,55 min.(A1)
Rt = 33,11 min.{B1)
LC-MS: 623.6 [M+1 ]' __.
